Sulfonamide Derivatives For The Treatment of Diseases

ABSTRACT

The invention relates to compounds of formula (1) 
     
       
         
         
             
             
         
       
     
     and to processes for the preparation of, intermediates used in the preparation of, compositions containing and the uses of, such derivatives. The compounds according to the present invention are useful in numerous diseases, disorders and conditions, in particular inflammatory, allergic and respiratory diseases, disorders and conditions.

This invention relates to β2 agonists of general formula:

in which R¹, R², n and Q¹ have the meanings indicated below, and toprocesses for the preparation of, compositions containing and the usesof such derivatives.

Adrenoceptors are members of the large G-protein coupled receptorsuper-family. The adrenoceptor subfamily is itself divided into the αand β subfamilies with the p sub-family being composed of at least 3receptor sub-types: β1, β2 and β3. These receptors exhibit differentialexpression patterns in tissues of various systems and organs of mammals.β2 adrenergic (β2) receptors are mainly expressed in smooth muscle cells(e.g. vascular, bronchial, uterine or intestinal smooth muscles),whereas β3 adrenergic receptors are mainly expressed in fat tissues(therefore β3 agonists could potentially be useful in the treatment ofobesity and diabetes) and β1 adrenergic receptors are mainly expressedin cardiac tissues (therefore β1 agonists are mainly used as cardiacstimulants).

The pathophysiology and treatments of airway diseases have beenextensively reviewed in the literature (for reference see Barnes, P. J.Chest, 1997, 111:2, pp 17S-26S and Bryan, S. A. et al, Expert Opinion oninvestigational drugs, 2000, 9:1, pp 2542) and therefore only a briefsummary will be included here to provide some background information.

Glucocorticosteroids, anti-leukotrienes, theophylline, cromones,anti-cholinergics and β2 agonists constitute drug classes that arecurrently used to treat allergic and non-allergic airways diseases suchas asthma and chronic obstructive airways disease (COPD). Treatmentguidelines for these diseases include both short and long acting inhaledβ2 agonists. Short acting, rapid onset β2 agonists are used for “rescue”bronchodilation, whereas, long-acting forms provide sustained relief andare used as maintenance therapy.

Bronchodilation is mediated via agonism of the β2 adrenoceptor expressedon airway smooth muscle cells, which results in relaxation and hencebronchodilation. Thus, as functional antagonists, β2 agonists canprevent and reverse the effects of all bronchoconstrictor substances,including leukotriene D4 (LTD4), acetylcholine, bradykinin,prostaglandins, histamine and endothelins. Because β2 receptors are sowidely distributed in the airway, β2 agonists may also affect othertypes of cells that play a role in asthma. For example, it has beenreported that β2 agonists may stabilize mast cells. The inhibition ofthe release of bronchoconstrictor substances may be how β2 agonistsblock the bronchoconstriction induced by allergens, exercise and coldair. Furthermore, β2 agonists inhibit cholinergic neurotransmission inthe human airway, which can result in reduced cholinergic-reflexbronchoconstriction.

In addition to the airways, it has also been established that β2adrenoceptors are also expressed in other organs and tissues and thus β2agonists, such as those described in the present invention, may haveapplication in the treatment of other diseases such as, but not limitedto those of the nervous system, premature labor, congestive heartfailure, depression, inflammatory and allergic skin diseases, psoriasis,proliferative skin diseases, glaucoma and in conditions where there isan advantage in lowering gastric acidity, particularly in gastric andpeptic ulceration.

However, numerous β2 agonists are limited in their use due to their lowselectivity or adverse side-effects driven by high systemic exposure andmainly mediated through action at β2 adrenoreceptors expressed outsidethe airways (muscle tremor, tachycardia, palpitations, restlessness).Therefore there is a need for improved agents in this class.

Accordingly, there is still a need for novel β2 agonists that would havean appropriate pharmacological profile, for example in terms of potency,pharmacokinetics or duration of action. In this context, the presentinvention relates to novel β2 agonists.

Various Sulfonamide derivatives have already been disclosed. Forexample, WO02066250 discloses compounds active as β3 agonist, selectiveover β2, of formula

wherein m may be 2, R₁ may be H, OH or NR₅SO₂R₅ (R₅ being H or C₁-C₆alkyl), Z may be a bond, R₂ may be H or C₁-C₆ alkyl, R₄ may be C₁-C₆alkyl, B may be phenyl, Y is C₁-C₆ alkyl and A may be phenyl.

WO02/000622 discloses selective β3 agonists of formula:

wherein R¹ may be phenyl substituted with hydroxy andalkylsulfonylamino, X₁ may be a bond, R² may be hydrogen, R³ is hydrogenor hydroxyalkyl, X₂ may be CH₂, X₃ is a bond, O or NH and R⁴ is cyclicgroup.

Other sulfonamide derivatives are also disclosed in U.S. Pat. No.5,776,983 as β3 agonists They are more specifically of formula:

wherein R¹ may be CH₃, R² may be OH, R⁶ may be H, R³ may be H or alkyl,R⁴ may be H, alkyl, R⁵ may be H, R^(5′) may be C(O)NR⁵R⁶′ wherein R⁶ andR⁶′ may be H or lower alkyl.

However, none of the above sulfonamide derivatives have shown a β2agonist activity and a pharmacological profile allowing them to be usedas efficient drugs in the treatment of the β2-mediated diseases and/orconditions, in particular allergic and non-allergic airways diseases orother diseases such as those previously cited.

The invention relates to the compounds of general formula (1):

wherein the (CH₂)_(n)—C(═O)Q¹ group is in the meta or para position, R¹and R² are independently selected from H and C₁-C₄ alkyl, n is 0, 1 or 2and Q¹ is a group selected from:

and a group *—NR⁸-Q²-A, wherein p is 1 or 2, Q² is a C₁-C₄ alkyleneoptionally substituted with OH, R⁸ is H or C₁-C₄ alkyl and A is pyridyloptionally substituted with OH, C₃-C₇ cycloalkyl optionally substitutedwith OH or a group

wherein R³, R⁴, R⁵, R⁶ and R⁷ are the same or different and are selectedfrom H, C₁-C₄ alkyl, OR⁹, SR⁹, halo, CN, CF₃, OCF₃, COOR⁹, SO₂NR⁹R¹⁰,CONR⁹R¹⁰, NR⁹R¹⁰, NHCOR¹⁰ and phenyl optionally substituted with 1 to 3groups selected from OR⁹, halo and C₁-C₄ alkyl;wherein R⁹ and R¹⁰ are the same or different and are selected from H orC₁-C₄ alkyl and the * represent the attachment point to the carbonylgroup;wherein the group Q¹ is substituted at least with one hydroxy group;or, if appropriate, their pharmaceutically acceptable salts and/orisomers, tautomers, solvates or isotopic variations thereof.

The compounds of formula (1) are agonists of the β2 receptors, that areparticularly useful for the treatment of β2-mediated diseases and/orconditions, by showing excellent potency, in particular whenadministered via the inhalation route.

In the here above general formula (1), C₁-C₄ alkyl and C₁-C₄ alkylenedenote a straight-chain or branched group containing 1, 2, 3 or 4 carbonatoms. This also applies if they carry substituents or occur assubstituents of other radicals, for example in O—(C₁-C₄)alkyl radicals,S—(C₁-C₄)alkyl radicals etc. . . . Examples of suitable (C₁-C₄)alkylradicals are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl . . . . Examples of suitable (C₁-C₄)alkoxyradicals are methoxy, ethoxy, n-propyloxy, isopropyloxy, n-butyloxy,iso-butyloxy, sec-butyloxy and tert-butyloxy . . . .

The C₃-C₇ cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl. Preferred C₃-C₇ cycloalkyl are substitutedwith OH.

Finally, halo denotes a halogen atom selected from the group consistingof fluoro, chloro, bromo and iodo in particular fluoro or chloro.

In the following, the free bond on the phenyl group such as in thestructure below,

means that the phenyl can be substituted in the meta or para position.

The compounds of the formula (1)

can be prepared using conventional procedures such as by the followingillustrative methods in which R¹, R², Q¹, and n are as previouslydefined for the compounds of the formula (1) unless otherwise stated.

The amide derivatives of the formula (1) may be prepared by coupling anacid of formula (2) or a salt thereof:

with an amine of formula NHR⁸-Q²-A (3),

The coupling is generally carried out in an excess of said amine as anacid receptor, with a conventional coupling agent (e.g.1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride orN,N′-dicyclohexylcarbodiimide), optionally in the presence of a catalyst(e.g. 1-hydroxybenzotriazole hydrate or 1-hydroxy-7-azabenzotriazole),and optionally in the presence of a tertiary amine base (e.g.N-methylmorpholine, triethylamine or diisopropylethylamine). Thereaction may be undertaken in a suitable solvent such as pyridine,dimethylformamide, tetrahydrofuran, dimethylsulfoxide, dichloromethaneor ethyl acetate, and at temperature comprised between 10° C. and 40° C.(room temperature) for a period of 1-24 hours.

Said amine (3), (3′) or (3″) is either commercially available or may beprepared by conventional methods well known to the one skilled in theart (e.g. reduction, oxidation, alkylation, transition metal-mediatedcoupling, protection, deprotection etc. . . . ) from commerciallyavailable material.

The acid of formula (2) may be prepared from the corresponding ester offormula (4):

wherein Ra is a suitable acid protecting group, preferably a(C₁-C₄)alkyl group, which includes, but is not limited to, methyl andethyl, according to any method well-known to the one skilled in the artto prepare an acid from an ester, without modifying the rest of themolecule. For example, the ester may be hydrolysed by treatment withaqueous acid or base (e.g. hydrogen chloride, potassium hydroxide,sodium hydroxide or lithium hydroxide), optionally in the presence of asolvent or mixture of solvents (e.g. water, propionitrile, 1,4-dioxan,tetrahydrofuran/water), at a temperature comprised between 20° C. and100° C., for a period of 1 to 40 hours.

The ester of formula (4) may be prepared by reaction of an amine offormula (5):

wherein Ra and n are as previously defined, with a bromide of formula(6):

In a typical procedure, the amine of formula (5) is reacted with abromide of formula (6) optionally in the presence of a solvent ormixture of solvents (e.g. dimethyl sulphoxide, toluene,N,N-dimethylformamide, propionitrile, acetonitrile), optionally in thepresence of a suitable base (e.g. triethylamine, diisopropylethylamine,potassium carbonate, potassium hydrogen carbonate) at a temperaturecomprised between 80° C. and 120° C., for 12 to 48 hours.

The bromide of formula (6) may be prepared according to the method of WO02/06258 (pg. 36, example 14a).

Alternatively, the ester of formula (4) where n=1 may be prepared fromthe bromide of formula (7):

In a typical procedure the bromide (7) is treated with a suitablepalladium catalyst (e.g. [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)) under an atmosphere of carbon monoxide using RaOH assolvent (e.g. MeOH, EtOH) at elevated temperature (100° C.) and pressure(up to 100 psi) to give the ester of formula (4).

The amine of formula (5), where R₁ is Me and R₂ is H, may be prepared aseither the (R) or (S) enantiomer from the corresponding protected amineof formula (8):

wherein Ra and n are as previously defined and Rb and Rc represent anysuitable substituents so that HNRbRc is a chiral amine (for example, Rbmay be hydrogen and Rc may be α-methylbenzyl), provided that the bondsbetween N and Rb and N and Rc can be easily cleaved to give the freeamine of formula (5) using standard methodology for cleaving nitrogenprotecting groups, such as those found in the text book T. W. GREENE,Protective Groups in Organic Synthesis, A. Wiley-IntersciencePublication, 1981.

The amine of formula (8) may be prepared as a single diastereomer byreaction of an amine of formula HNRbRc with a ketone of formula (9):

wherein Ra, Rb, Rc and n are as previously defined.

In a typical procedure, the reaction of the ketone of formula (9) withthe amine of formula HNRbRc leads to a chiral intermediate which is inturn reduced by a suitable reducing agent (e.g. sodium cyanoborohydrideof formula NaCNBH₃ or sodium triacetoxyborohydride of formulaNa(OAc)₃BH) optionally in the presence of a drying agent (e.g. molecularsieves, magnesium sulfate) and optionally in the presence of an acidcatalyst (e.g. acetic acid) to give the amine of formula (8) as amixture of diastereomers. The reaction is generally done in a solventsuch as tetrahydrofuran or dichloromethane at a temperature comprisedbetween 20° C. and 80° C. for 3 to 72 hours. The resulting product isthen converted to the hydrochloride salt and selectively crystallisedfrom a suitable solvent or mixture of solvents (e.g. isopropanol,ethanol, methanol, diisopropyl ether or diisopropyl ether/methanol) togive (8) as a single diastereomer.

The ketone of formula (9) where n=1 may be prepared by palladiummediated coupling of an aryl halide of formula (10):

wherein Ra is as previously defined and Hal represents an halogen atom,which includes, but is not limited to bromo and iodo, with an enolate orenolate equivalent.

In a typical procedure, the aryl halide of formula (10) is reacted witha tin enolate generated in-situ by treatment of isoprenyl acetate withtri-n-butyltin methoxide of formula Bu₃SnOMe in the presence of asuitable palladium catalyst (palladium acetate/tri-ortho-tolylphosphineof formula Pd(OAc)₂/P(o-Tol)₃) in a non-polar solvent (e.g. toluene,benzene, hexane). Preferably, the reaction is carried out at atemperature comprised between 80° C. and 110° C. for 6 to 16 hours.

The aryl halide of formula (10) may be obtained by esterification of thecorresponding acid of formula (11):

wherein Hal is as previously defined, according to any method well-knownto the one skilled in the art to prepare an ester from an acid, withoutmodifying the rest of the molecule.

In a typical procedure, the acid of formula (11) is reacted with analcoholic solvent of formula RaOH, wherein Ra is as previously defined,in the presence of an acid such as hydrogen chloride at a temperaturebetween 10° C. and 40° C. (room temperature) for 8 to 16 hours.

The acid of formula (11) is a commercial product.

The amine of formula (5), where R₁═R₂=alkyl, may be prepared accordingto the following scheme:

wherein R¹, R² and Ra are as previously defined.

In a typical procedure, the ester of formula (12) is reacted with an“activated” alkyl (organometallic alkyl such as R²MgBr, R²MgCl or R²Li)to give the corresponding tertiary alcohol of formula (13) using themethod described above.

Said tertiary alcohol of formula (13) is then treated with an alkylnitrile (e.g. acetonitrile, chloroacetonitrile) in the presence of anacid (e.g. sulphuric acid, acetic acid) to give a protected intermediatewhich is in turn cleaved using standard methodology for cleavingnitrogen protecting group such as those mentioned in textbooks. Theresulting aminoacid is then esterified using the method described hereinto give the amine of formula (5).

Alternatively, the amine of formula (5), where R¹═R²═C₁-C₄ alkyl andn=0, may be prepared according to the following scheme:

wherein R¹, R² and Ra are as previously defined.

In a typical procedure, the ester of formula (14) is reacted with an“activated” alkyl (organometallic alkyl such as R²MgBr, R²MgCl or R²Li)to give the corresponding tertiary alcohol of formula (15) using themethod described above.

Said tertiary alcohol of formula (15) is then treated with an alkylnitrile (e.g. acetonitrile, chloroacetonitrile) in the presence of anacid (e.g. sulphuric acid, acetic acid) to give a protected intermediatewhich is in turn cleaved using standard methodology for cleavingnitrogen protecting group such as those mentioned in textbooks to givethe bromo amine (16).

The resulting bromo amine (16) is treated with a suitable palladiumcatalyst (e.g.[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)) under anatmosphere of carbon monoxide using RaOH as solvent (e.g. MeOH, EtOH) atelevated temperature (100° C.) and pressure (100 psi) to give the esterof formula (5).

The ketone of formula (9) where n=2 may be prepared by reduction of analkene of formula (17):

In a typical procedure, a solution of the olefin of formula (17) in asuitable solvent (e.g. methanol, ethanol, ethyl acetate) is treated witha palladium catalyst (e.g. 10% palladium on charcoal) and stirred underan atmosphere of hydrogen, optionally at elevated pressure (e.g. 60psi), at temperature between room temperature and 60° C. for 8-24 hours.

The alkene of formula (17) may be prepared by a palladium mediatedcoupling of an activated olefin with an aryl halide of formula (18):

In a typical procedure, the aryl halide (18) is coupled with a vinylester (e.g. methyl acrylate) in the presence of a suitable palladiumcatalyst (e.g. tetrakis(triphenylphosphine)palladium(0) of formulaPd(PPh₃)₄, palladium acetate/tri-ortho-tolylphosphine of formulaPd(OAc)₂/P(o-tol)₃ or (diphenylphosphino)ferrocenyl palladium chlorideof formula dppfPdCl₂) in a suitable solvent (e.g. acetonitrile,N,N-dimethylformamide, toluene), optionally in the presence of a basesuch as triethylamine at a temperature between 40° C. and 110° C. for 8to 24 hours.

The ketone of formula (18) is a commercial product.

The amine of formula (5), where R¹ and R² are both H, may be preparedaccording to the following scheme:

wherein R¹, R² and Ra are as previously defined.

In a typical procedure, the acid of formula (19) is preferentiallyreduced to the corresponding alcohol (20) in the presence of the ester.This may be performed by formation of the acyl imidazole or mixedanhydride and subsequent reduction with sodium borohydride or anothersuitable reducing agent.

Said primary alcohol of formula (20) is then converted into a leavinggroup such as mesylate, tosylate, bromide or iodide and displaced withappropriate amine nucleophile. The preferred nucleophile is azide ionwhich can then be reduced to the primary amine via hydrogenation ortriphenylphosphine. Alternative nucleophiles could include ammonia oralkylamines such as benzylamine or allylamine and subsequent cleavage ofthe alkyl group to furnish the amine.

In a typical procedure, the compounds of formula (I) wherein R¹ and R²are both methyl and n is 1, may be prepared by reacting a compound offormula (21)

where X is H, K, Na, Li and potentially an organic amine base or othermetal salt, with a suitable amine of formula NHR⁸-Q²-A (3)

in the presence of a conventional coupling agent such as1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride ordicyclohexylcarbodiimide in a suitable solvent such as pyridinedimethylformamide and dimethylacetamide optionally in the presence of anorganic base (such as Hunig's base) and an additive (such as1-hydroxybenzotriazole) in order to obtain a compound of formula (1):

wherein R¹ and R² are methyl and n is 1.

Said compound of formula (21) may be obtained by hydrogenation of acompound of formula (22)

wherein X is H, Na, Li or K and an potentially an organic amine or othermetal salts, in the presence of a suitable solvent such as methanol,IPA, THF and water and in the presence of a suitable catalyst such aspalladium hydroxide on carbon or palladium on carbon.

Said compound of formula (22) may be obtained by reacting a compound offormula (23)

with M—OH wherein M is selected from Na, K or Li, optionally in thepresence of a suitable solvent such as propionitrile, tetrahydrofuran ordioxane, preferably propionitrile.

Said compound of formula (23) may be obtained by deprotecting a compoundof formula (24)

using a deprotecting agent such as tetrabutylammonium fluoride, HF, ortriethylamine trihydrofluoride in the presence of a suitable solventsuch as propionitrile.

Said compound of formula (24) may be obtained by reacting a compound offormula

with a compound of formula

in the presence of a suitable solvent such as propionitrile, THF,toluene, ethyl acetate, acetonitrile, propionitrile, dioxane, DMF, DMSO,and optionally in the presence of a base such as sodium hydrogencarbonate, potassium hydrogen carbonate, Hunig's base or triethylamine,at a temperature between 50° C. and 150° C. for 12 to 36 hours.

For some of the steps of the here above described process of preparationof the compounds of formula (1), it may be necessary to protectpotential reactive functions that are not wished to react, and to cleavesaid protecting groups in consequence. In such a case, any compatibleprotecting radical can be used. In particular methods of protection anddeprotection such as those described by T. W. GREENE (Protective Groupsin Organic Synthesis, A. Wiley-Interscience Publication, 1981) or by P.J. Kocienski (Protecting groups, Georg Thieme Verlag, 1994), can beused.

All of the above reactions and the preparations of novel startingmaterials used in the preceding methods are conventional and appropriatereagents and reaction conditions for their performance or preparation aswell as procedures for isolating the desired products will be well-knownto those skilled in the art with reference to literature precedents andthe examples and preparations hereto.

Also, the compounds of formula (1) as well as intermediate for thepreparation thereof can be purified according to various well-knownmethods, such as for example crystallization or chromatography.

Preferably Q² is —CH₂—, —(CH₂)₂—, —(CH₂)₃—, —(C(CH₃)₂)—, —(CH₂)₄— or—(CH(CH₂OH))—.

Preferably, Q¹ is

wherein one of R₃, R₄, R₅ and R₆ is OH and the others are H.

Preferably Q¹ is

wherein one of R₃, R₄, R₅ and R₆ is OH and the others are H.

Preferably Q¹ is a group *—NR⁸-Q²-A, wherein R⁸ is H, CH₃ or CH₂CH₃, Q²is a C₁-C₄ alkylene and A is naphtyl substituted by one hydroxy.

Preferably, Q¹ is a group *—NR⁸-Q²-A, wherein R⁸ is H, CH₃ or CH₂CH₃, Q²is a C₁-C₄ alkylene and A is a group

wherein one of R³, R⁴, R⁵, R⁶ and R⁷ is OH and the others are the sameor different and are selected from H, C₁-C₄ alkyl, OR⁹, SR⁹, halo, CF₃,OCF₃, SO₂NR⁹R¹⁰, CONR⁹R¹⁰, NR⁹R¹⁰, NHCOR¹⁰, provided at least 2 of R³ toR⁷ are equal to H;

wherein R⁹ and R¹⁰ are the same or different and are selected from H orC₁-C₄ alkyl.

More preferably, Q¹ is a group *—NH-Q²-A, wherein Q² is a —CH₂—,—(CH₂)₂—, —(CH₂)₄—, —CH₂—C(CH₃)₂)— preferably —CH₂—, and A is a group

wherein one of R³, R⁴, R⁵, R⁵ and R⁷ is OH and the others are the sameor different and are selected from H, OH, CH₃, OCH₂—CH₃, SCH₃, halo,CF₃, OCF₃, provided at least 2 of R³ to R⁷ are equal to H.

Preferably, Q¹ is a group *—NR⁸-Q²-A, wherein R⁸ is H, CH₃ or CH₂CH₃, Q²is a C₁-C₄ alkylene and A is a group

wherein one of R³, R⁴, R⁵, R⁶ and R⁷ is phenyl substituted by OH and theothers are H.

In the above groups of compounds, the following substituents areparticularly preferred:

R¹ is H or C₁-C₄ alkyl and R² is C₁-C₄ alkyl. More preferably, R¹ is Hor CH₃ and R² is CH₃.

n is 0 or 1. More preferably n is 1.

R¹ is H and R² is CH₃ and n is 1.

R¹ is CH₃, R² is CH₃ and n is 1.

Particularly preferred are the compounds of the formula (1) as describedin the Examples section hereafter, i.e.:

-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-(4-hydroxy-3-methoxybenzyl)acetamide;-   N-[(4′-Hydroxybiphenyl-4-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-(4-Chloro-2-hydroxybenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-(4-Hydroxy-3,5-dimethylbenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(2-hydroxy-1-naphthyl)methyl]acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(6-hydroxy-2-naphthyl)methyl]acetamide;-   N-[(4′-Hydroxybiphenyl-3-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-[(3′-Hydroxybiphenyl-3-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[2-(4-hydroxyphenyl)-2-methylpropyl]acetamide;-   N-(3,5-Dichloro-2-hydroxybenzyl)-N-ethyl-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(6-hydroxy-1-naphthyl)methyl]-N-methylacetamide;-   N-[(2′-Hydroxybiphenyl-3-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-(2-Hydroxy-5-{(1R)-1-hydroxy-2-[(2-{3-[2-(6-hydroxy-3,4-dihydroisoquinolin-2(1H)-yl)-2-oxoethyl]phenyl}-1,1-dimethylethyl)amino]ethyl}phenyl)methanesulfonamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[4-(4-hydroxyphenyl)butyl]acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[2-(4-hydroxyphenyl)ethyl]acetamide;-   N-(2-Chloro-4-hydroxybenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide-   N-(3,5-Dichloro-4-hydroxybenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-(2,3-Dichloro-4-hydroxybenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(4-hydroxy-1-naphthyl)methyl]acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[3-hydroxy-5-(trifluoromethyl)benzyl]acetamide;-   N-(2-Chloro-4-hydroxybenzyl)-N-ethyl-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-(2-Chloro-4-hydroxybenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-methylacetamide;-   N-(3-Fluoro-5-hydroxybenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-methylacetamide;-   N-[(2′-Hydroxybiphenyl-2-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-[(3′-Hydroxybiphenyl-2-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-(4-Hydroxy-2,6-dimethylbenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-(2-Hydroxy-5-{(1R)-1-hydroxy-2-[(2-{3-[2-(7-hydroxy-3,4-dihydroisoquinolin-2(1H)-yl)-2-oxoethyl]phenyl}-1,1-dimethylethyl)amino]ethyl}phenyl)methanesulfonamide;-   N-(2-Hydroxy-5-{(1R)-1-hydroxy-2-[(2-{3-[2-(5-hydroxy-3,4-dihydroisoquinolin-2(1H)-yl)-2-oxoethyl]phenyl}-1,1-dimethylethyl)amino]ethyl}phenyl)methanesulfonamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(1R)-2-hydroxy-1-phenylethyl]acetamide;-   2-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(1S)-2-hydroxy-1-phenylethyl]acetamide;-   N-[(3′-Hydroxybiphenyl-4-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-[(2′-Hydroxybiphenyl-4-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide;-   N-[(4′-Hydroxybiphenyl-4-yl)methyl]-3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}benzamide;-   3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}-N-[2-(4-hydroxyphenyl)-2-methylpropyl]benzamide;-   N-[(4′-Hydroxybiphenyl-3-yl)methyl]-3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}benzamide;-   N-[2-(4-Hydroxy-2,5-dimethylphenyl)ethyl]-3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}benzamide;-   N-[2-(4-Hydroxy-2,3-dimethylphenyl)ethyl]-3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}benzamide;    and,-   3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}-N-[2-(4-hydroxy-3-methylphenyl)ethyl]benzamide.

According to one aspect of the present invention, the compounds offormula (I) wherein the (CH₂)_(n)—C(═O)Q¹ group is in the meta positionare generally preferred.

Pharmaceutically acceptable salts of the compounds of formula (1)include the acid addition and base salts thereof.

Suitable acid addition salts are formed from acids which form non-toxicsalts. Examples include the acetate, aspartate, benzoate, besylate,bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, citrate,edisylate, esylate, formate, fumarate, gluceptate, gluconate,glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride,hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate,maleate, malonate, mesylate, methylsulphate, naphthylate, 2-napsylate,nicotinate, nitrate, orotate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate, stearate,succinate, tartrate, tosylate and trifluoroacetate and xinafoate salts.

Suitable base salts are formed from bases which form non-toxic salts.Examples include the aluminium, arginine, benzathine, calcium, choline,diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine,potassium, sodium, tromethamine and zinc salts.

Hemisalts of acids and bases may also be formed, for example,hemisulphate and hemicalcium salts.

For a review on suitable salts, see “Handbook of Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH,Weinheim, Germany, 2002).

Pharmaceutically acceptable salts of compounds of formula (1) may beprepared by one or more of three methods:

-   (i) by reacting the compound of formula (1) with the desired acid or    base;-   (ii) by removing an acid- or base-labile protecting group from a    suitable precursor of the compound of formula (1) or by ring-opening    a suitable cyclic precursor, for example, a lactone or lactam, using    the desired acid or base; or-   (iii) by converting one salt of the compound of formula (1) to    another by reaction with an appropriate acid or base or by means of    a suitable ion exchange column.

All three reactions are typically carried out in solution. The resultingsalt may precipitate out and be collected by filtration or may berecovered by evaporation of the solvent. The degree of ionisation in theresulting salt may vary from completely ionised to almost non-ionised.

The compounds of the invention may exist in both unsolvated and solvatedforms. The term ‘solvate’ is used herein to describe a molecular complexcomprising the compound of the invention and a stoichiometric amount ofone or more pharmaceutically acceptable solvent molecules, for example,ethanol. The term ‘hydrate’ is employed when said solvent is water.

Included within the scope of the invention are complexes such asclathrates, drug-host inclusion complexes wherein, in contrast to theaforementioned solvates, the drug and host are present in stoichiometricor non-stoichiometric amounts. Also included are complexes of the drugcontaining two or more organic and/or inorganic components which may bein stoichiometric or non-stoichiometric amounts. The resulting complexesmay be ionised, partially ionised, or non-ionised. For a review of suchcomplexes, see J Pharm Sci, 64 (8), 1269-1288 by Haleblian (August1975).

Hereinafter all references to compounds of formula (1) includereferences to salts, solvates and complexes thereof and to solvates andcomplexes of salts thereof.

The compounds of the invention include compounds of formula (1) ashereinbefore defined, including all polymorphs and crystal habitsthereof, prodrugs and isomers thereof (including optical, geometric andtautomeric isomers) as hereinafter defined and isotopically-labeledcompounds of formula (1).

As indicated, so-called ‘pro-drugs’ of the compounds of formula (1) arealso within the scope of the invention. Thus certain derivatives ofcompounds of formula (1) which may have little or no pharmacologicalactivity themselves can, when administered into or onto the body, beconverted into compounds of formula (1) having the desired activity, forexample, by hydrolytic cleavage. Such derivatives are referred to as‘prodrugs’. Further information on the use of prodrugs may be found in‘Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T.Higuchi and W. Stella) and ‘Bioreversible Carriers in Drug Design’,Pergamon Press, 1987 (ed. E. B Roche, American PharmaceuticalAssociation).

Prodrugs in accordance with the invention can, for example, be producedby replacing appropriate functionalities present in the compounds offormula (1) with certain moieties known to those skilled in the art as‘pro-moieties’ as described, for example, in “Design of Prodrugs” by H.Bundgaard (Elsevier, 1985).

Some examples of prodrugs in accordance with the invention include:

-   (i) where the compound of formula (1) contains a carboxylic acid    functionality (—COOH), an ester thereof, for example, a compound    wherein the hydrogen of the carboxylic acid functionality of the    compound of formula (1) is replaced by (C₁-C₈)alkyl;-   (ii) where the compound of formula (1) contains an alcohol    functionality (—OH), an ether thereof, for example, a compound    wherein the hydrogen of the alcohol functionality of the compound of    formula (1) is replaced by (C₁-C₆)alkanoyloxymethyl; and-   (iii) where the compound of formula (1) contains a primary or    secondary amino functionality (—NH₂ or —NHR where R≠H), an amide    thereof, for example, a compound wherein, as the case may be, one or    both hydrogens of the amino functionality of the compound of    formula (1) is/are replaced by (C₁-C₁₀)alkanoyl.

Further examples of replacement groups in accordance with the foregoingexamples and examples of other prodrug types may be found in theaforementioned references.

Moreover, certain compounds of formula (1) may themselves act asprodrugs of other compounds of formula (1).

Also included within the scope of the invention are metabolites ofcompounds of formula (1), that is, compounds formed in vivo uponadministration of the drug. Some examples of metabolites in accordancewith the invention include

-   (i) where the compound of formula (1) contains a methyl group, an    hydroxymethyl derivative thereof (—CH₃→—CH₂OH):-   (ii) where the compound of formula (1) contains an alkoxy group, an    hydroxy derivative thereof (—OR→—OH);-   (iii) where the compound of formula (1) contains a tertiary amino    group, a secondary amino derivative thereof (—NR¹R²→—NHR¹ or —NHR²);-   (iv) where the compound of formula (1) contains a secondary amino    group, a primary derivative thereof (—NHR¹→—NH₂);-   (v) where the compound of formula (1) contains a phenyl moiety, a    phenol derivative thereof (-Ph→-PhOH); and-   (vi) where the compound of formula (1) contains an amide group, a    carboxylic acid derivative thereof (—CONH₂→COOH).

Compounds of formula (1) containing one or more asymmetric carbon atomscan exist as two or more stereoisomers. Where a compound of formula (1)contains an alkenyl or alkenylene group, geometric cis/trans (or Z/E)isomers are possible. Where structural isomers are interconvertible viaa low energy barrier, tautomeric isomerism (‘tautomerism’) can occur.This can take the form of proton tautomerism in compounds of formula (1)containing, for example, an imino, keto, or oxime group, or so-calledvalence tautomerism in compounds which contain an aromatic moiety. Itfollows that a single compound may exhibit more than one type ofisomerism.

Included within the scope of the present invention are allstereoisomers, geometric isomers and tautomeric forms of the compoundsof formula (1), including compounds exhibiting more than one type ofisomerism, and mixtures of one or more thereof. Also included are acidaddition or base salts wherein the counterion is optically active, forexample, d-lactate or l-lysine, or racemic, for example, dl-tartrate ordl-arginine.

Cis/trans isomers may be separated by conventional techniques well knownto those skilled in the art, for example, chromatography and fractionalcrystallisation.

Conventional techniques for the preparation/isolation of individualenantiomers include chiral synthesis from a suitable optically pureprecursor or resolution of the racemate (or the racemate of a salt orderivative) using, for example, chiral high pressure liquidchromatography (HPLC).

Alternatively, the racemate (or a racemic precursor) may be reacted witha suitable optically active compound, for example, an alcohol, or, inthe case where the compound of formula (1) contains an acidic or basicmoiety, an acid or base such as tartaric acid or 1-phenylethylamine. Theresulting diastereomeric mixture may be separated by chromatographyand/or fractional crystallization and one or both of thediastereoisomers converted to the corresponding pure enantiomer(s) bymeans well known to a skilled person.

Chiral compounds of the invention (and chiral precursors thereof) may beobtained in enantiomerically-enriched form using chromatography,typically HPLC, on an asymmetric resin with a mobile phase consisting ofa hydrocarbon, typically heptane or hexane, containing from 0 to 50% byvolume of isopropanol, typically from 2% to 20%, and from 0 to 5% byvolume of an alkylamine, typically 0.1% diethylamine. Concentration ofthe eluate affords the enriched mixture.

Stereoisomeric conglomerates may be separated by conventional techniquesknown to those skilled in the art—see, for example, “Stereochemistry ofOrganic Compounds” by E. L. Eliel (Wiley, New York, 1994).

According to one aspect of the present invention, the (R,R)-stereoisomerof the formula below, wherein R¹ is hydrogen and R² is C₁-C₄ alkyl,preferably methyl, and n and Q¹ are as defined above, is generallypreferred:

The present invention includes all pharmaceutically acceptableisotopically-labelled compounds of formula (1) wherein one or more atomsare replaced by atoms having the same atomic number, but an atomic massor mass number different from the atomic mass or mass number whichpredominates in nature.

Examples of isotopes suitable for inclusion in the compounds of theinvention include isotopes of hydrogen, such as ²H and ³H, carbon, suchas ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶Cl, fluorine, such as ¹⁸F,iodine, such as ¹²³I and ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N, oxygen,such as ¹⁵O, ¹⁷O and ¹⁸O, phosphorus, such as ³²P, and sulphur, such as³⁵S.

Certain isotopically-labelled compounds of formula (1), for example,those incorporating a radioactive isotope, are useful in drug and/orsubstrate tissue distribution studies. The radioactive isotopes tritium,i.e. ³H, and carbon-14, i.e. ¹⁴C, are particularly useful for thispurpose in view of their ease of incorporation and ready means ofdetection.

Substitution with heavier isotopes such as deuterium, i.e. ²H, mayafford certain therapeutic advantages resulting from greater metabolicstability, for example, increased in vivo half-life or reduced dosagerequirements, and hence may be preferred in some circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, can be useful in Positron Emission Topography (PET) studies forexamining substrate receptor occupancy.

Isotopically-labeled compounds of formula (1) can generally be preparedby conventional techniques known to those skilled in the art or byprocesses analogous to those described in the accompanying Examples andPreparations using an appropriate isotopically-labeled reagents in placeof the non-labeled reagent previously employed.

Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d₆-acetone, d₆-DMSO.

The compounds of formula (1), their pharmaceutically acceptable saltsand/or derived forms, are valuable pharmaceutically active compounds,which are suitable for the therapy and prophylaxis of numerous disordersin which the O₂ receptor is involved or in which agonism of thisreceptor may induce benefit, in particular the allergic and non-allergicairways diseases but also in the treatment of other diseases such as,but not limited to those of the nervous system, premature labor,congestive heart failure, depression, inflammatory and allergic skindiseases, psoriasis, proliferative skin diseases, glaucoma and inconditions where there is an advantage in lowering gastric acidity,particularly in gastric and peptic ulceration.

Compounds of the invention intended for pharmaceutical use may beadministered as crystalline or amorphous products. They may be obtained,for example, as solid plugs, powders, or films by methods such asprecipitation, crystallization, freeze drying, spray drying, orevaporative drying. Microwave or radio frequency drying may be used forthis purpose.

They may be administered alone or in combination with one or more othercompounds of the invention or in combination with one or more otherdrugs (or as any combination thereof). Generally, they will beadministered as a formulation in association with one or morepharmaceutically acceptable excipients. The term “excipient” is usedherein to describe any ingredient other than the compound(s) of theinvention. The choice of excipient will to a large extent depend onfactors such as the particular mode of administration, the_effect of theexcipient on solubility and stability, and the nature of the dosageform.

Pharmaceutical compositions suitable for the delivery of compounds ofthe present invention and methods for their preparation will be readilyapparent to those skilled in the art. Such compositions and methods fortheir preparation may be found, for example, in ‘Remington'sPharmaceutical Sciences’, 19th Edition (Mack Publishing Company, 1995).

The compounds of the invention may also be administered directly intothe blood stream, into muscle, or into an internal organ. Suitable meansfor parenteral administration include intravenous, intraarterial,intraperitoneal, intrathecal, intraventricular, intraurethral,intrasternal, intracranial, intramuscular and subcutaneous. Suitabledevices for parenteral administration include needle (includingmicroneedle) injectors, needle-free injectors and infusion techniques.

Parenteral formulations are typically aqueous solutions which maycontain excipients such as salts, carbohydrates and buffering agents(preferably to a pH of from 3 to 9), but, for some applications, theymay be more suitably formulated as a sterile non-aqueous solution or asa dried form to be used in conjunction with a suitable vehicle such assterile, pyrogen-free water.

The preparation of parenteral formulations under sterile conditions, forexample, by lyophilisation, may readily be accomplished using standardpharmaceutical techniques well known to those skilled in the art.

The solubility of compounds of formula (1) used in the preparation ofparenteral solutions may be increased by the use of appropriateformulation techniques, such as the incorporation ofsolubility-enhancing agents.

Formulations for parenteral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease. Thus compounds of the invention may be formulated as a solid,semi-solid, or thixotropic liquid for administration as an implanteddepot providing modified release of the active compound. Examples ofsuch formulations include drug-coated stents andpoly(dl-lactic-coglycolic)acid (PGLA) microspheres.

The compounds of the invention may also be administered topically to theskin or mucosa, that is, dermally or transdermally. Typical formulationsfor this purpose include gels, hydrogels, lotions, solutions, creams,ointments, dusting powders, dressings, foams, films, skin patches,wafers, implants, sponges, fibres, bandages and microemulsions.Liposomes may also be used. Typical carriers include alcohol, water,mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethyleneglycol and propylene glycol. Penetration enhancers may beincorporated—see, for example, J Pharm Sci, 88 (10), 955-958 by Finninand Morgan (October 1999).

Other means of topical administration include delivery byelectroporation, iontophoresis, phonophoresis, sonophoresis andmicroneedle or needle-free (e.g. Powderject™, Bioject™, etc.) injection.

Formulations for topical administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

The compounds of the invention can also be administered intranasally orby inhalation, typically in the form of a dry powder (either alone, as amixture, for example, in a dry blend with lactose, or as a mixedcomponent particle, for example, mixed with phospholipids, such asphosphatidylcholine) from a dry powder inhaler or as an aerosol sprayfrom a pressurised container, pump, spray, atomiser (preferably anatomiser using electrohydrodynamics to produce a fine mist), ornebuliser, with or without the use of a suitable propellant, such as1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. Forintranasal use, the powder may comprise a bioadhesive agent, forexample, chitosan or cyclodextrin.

The pressurised container, pump, spray, atomizer, or nebuliser containsa solution or suspension of the compound(s) of the invention comprising,for example, ethanol, aqueous ethanol, or a suitable alternative agentfor dispersing, solubilising, or extending release of the active, apropellant(s) as solvent and an optional surfactant, such as sorbitantrioleate, oleic acid, or an oligolactic acid.

Prior to use in a dry powder or suspension formulation, the drug productis micronised to a size suitable for delivery by inhalation (typicallyless than 5 microns). This may be achieved by any appropriatecomminuting method, such as spiral jet milling, fluid bed jet milling,supercritical fluid processing to form nanoparticles, high pressurehomogenisation, or spray drying.

Capsules (made, for example, from gelatin orhydroxypropylmethylcellulose), blisters and cartridges for use in aninhaler or insufflator may be formulated to contain a powder mix of thecompound of the invention, a suitable powder base such as lactose orstarch and a performance modifier such as l-leucine, mannitol, ormagnesium stearate. The lactose may be anhydrous or in the form of themonohydrate, preferably the latter. Other suitable excipients includedextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose andtrehalose.

A suitable solution formulation for use in an atomiser usingelectrohydrodynamics to produce a fine mist may contain from 1 μg to 20mg of the compound of the invention per actuation and the actuationvolume may vary from 1 μl to 100 μl. A typical formulation may comprisea compound of formula (1), propylene glycol, sterile water, ethanol andsodium chloride. Alternative solvents which may be used instead ofpropylene glycol include glycerol and polyethylene glycol.

Suitable flavours, such as menthol and levomenthol, or sweeteners, suchas saccharin or saccharin sodium, may be added to those formulations ofthe invention intended for inhaled/intranasal administration.

Formulations for inhaled/intranasal administration may be formulated tobe immediate and/or modified release using, for example, PGLA. Modifiedrelease formulations include delayed-, sustained-, pulsed-, controlled-,targeted and programmed release.

In the case of dry powder inhalers and aerosols, the dosage unit isdetermined by means of a valve which delivers a metered amount. Units inaccordance with the invention are typically arranged to administer ametered dose or “puff” containing from 0.001 mg to 10 mg of the compoundof formula (1). The overall daily dose will typically be in the range0.001 mg to 40 mg which may be administered in a single dose or, moreusually, as divided doses throughout the day.

The compounds of formula (1) are particularly suitable for anadministration by inhalation

The compounds of the invention may be administered rectally orvaginally, for example, in the form of a suppository, pessary, or enema.Cocoa butter is a traditional suppository base, but various alternativesmay be used as appropriate.

Formulations for rectal/vaginal administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

The compounds of the invention may also be administered directly to theeye or ear, typically in the form of drops of a micronised suspension orsolution in isotonic, pH-adjusted, sterile saline. Other formulationssuitable for ocular and aural administration include ointments,biodegradable (e.g. absorbable gel sponges, collagen) andnon-biodegradable (e.g. silicone) implants, wafers, lenses andparticulate or vesicular systems, such as niosomes or liposomes. Apolymer such as crossed-linked polyacrylic acid, polyvinylalcohol,hyaluronic acid, a cellulosic polymer, for example,hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose, or a heteropolysaccharide polymer, for example, gelan gum,may be incorporated together with a preservative, such as benzalkoniumchloride. Such formulations may also be delivered by iontophoresis.

Formulations for ocular/aural administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted, or programmedrelease.

The compounds of the invention may be combined with solublemacromolecular entities, such as cyclodextrin and suitable derivativesthereof or polyethylene glycol-containing polymers, in order to improvetheir solubility, dissolution rate, taste-masking, bioavailabilityand/or stability for use in any of the aforementioned modes ofadministration.

Drug-cyclodextrin complexes, for example, are found to be generallyuseful for most dosage forms and administration routes. Both inclusionand non-inclusion complexes may be used. As an alternative to directcomplexation with the drug, the cyclodextrin may be used as an auxiliaryadditive, i.e. as a carrier, diluent, or solubiliser. Most commonly usedfor these purposes are alpha-, beta- and gamma-cyclodextrins, examplesof which may be found in International Patent Applications Nos. WO91/11172, WO 94/02518 and WO 98/55148.

Inasmuch as it may desirable to administer a combination of activecompounds, for example, for the purpose of treating a particular diseaseor condition, it is within the scope of the present invention that twoor more pharmaceutical compositions, at least one of which contains acompound in accordance with the invention, may conveniently be combinedin the form of a kit suitable for coadministration of the compositions.

Thus the kit of the invention comprises two or more separatepharmaceutical compositions, at least one of which contains a compoundof formula (1) in accordance with the invention, and means forseparately retaining said compositions, such as a container, dividedbottle, or divided foil packet. An example of such a kit is the familiarblister pack used for the packaging of tablets, capsules and the like.

The kit of the invention is particularly suitable for administeringdifferent dosage forms, for example parenteral, for administering theseparate compositions at different dosage intervals, or for titratingthe separate compositions against one another. To assist compliance, thekit typically comprises directions for administration and may beprovided with a so-called memory aid.

For administration to human patients, the total daily dose of thecompounds of the invention is typically in the range 0.001 mg to 5000 mgdepending, of course, on the mode of administration. For example, anintravenous daily dose may only require from 0.001 mg to 40 mg. Thetotal daily dose may be administered in single or divided doses and may,at the physician's discretion, fall outside of the typical range givenherein.

These dosages are based on an average human subject having a weight ofabout 65 kg to 70 kg. The physician will readily be able to determinedoses for subjects whose weight falls outside this range, such asinfants and the elderly.

For the avoidance of doubt, references herein to “treatment” includereferences to curative, palliative and prophylactic treatment.

According to another embodiment of the present invention, the compoundsof the formula (1), or pharmaceutically acceptable salts, derived formsor compositions thereof, can also be used as a combination with one ormore additional therapeutic agents to be co-administered to a patient toobtain some particularly desired therapeutic end result such as thetreatment of pathophysiologically-relevant disease processes including,but not limited to (i) bronchoconstriction, (ii) inflammation, (iii)allergy, (iv) tissue destruction, (v) signs and symptoms such asbreathlessness, cough. The second and more additional therapeutic agentsmay also be a compound of the formula (1), or a pharmaceuticallyacceptable salt, derived forms or compositions thereof, or one or moreβ2 agonists known in the art. More typically, the second and moretherapeutic agents will be selected from a different class oftherapeutic agents.

As used herein, the terms “co-administration”, “co-administered” and “incombination with”, referring to the compounds of formula (1) and one ormore other therapeutic agents, is intended to mean, and does refer toand include the following:

-   -   simultaneous administration of such combination of compound(s)        of formula (1) and therapeutic agent(s) to a patient in need of        treatment, when such components are formulated together into a        single dosage form which releases said components at        substantially the same time to said patient,    -   substantially simultaneous administration of such combination of        compound(s) of formula (1) and therapeutic agent(s) to a patient        in need of treatment, when such components are formulated apart        from each other into separate dosage forms which are taken at        substantially the same time by said patient, whereupon said        components are released at substantially the same time to said        patient,    -   sequential administration of such combination compound(s) of        formula (1) and therapeutic agent(s) to a patient in need of        treatment, when such components are formulated apart from each        other into separate dosage forms which are taken at consecutive        times by said patient with a significant time interval between        each administration, whereupon said components are released at        substantially different times to said patient; and    -   sequential administration of such combination of compound(s) of        formula (1) and therapeutic agent(s) to a patient in need of        treatment, when such components are formulated together into a        single dosage form which releases said components in a        controlled manner whereupon they are concurrently,        consecutively, and/or overlapingly administered at the same        and/or different times by said patient,        where each part may be administered by either the same or        different route.

Suitable examples of other therapeutic agents which may be used incombination with the compound(s) of formula (1), or pharmaceuticallyacceptable salts, derived forms or compositions thereof, include, butare by no means limited to:

-   (a) 5-Lipoxygenase (5-LO) inhibitors or 5-lipoxygenase activating    protein (FLAP) antagonists,-   (b) Leukotriene antagonists (LTRAs) including antagonists of LTB₄,    LTC₄, LTD₄, and LTE₄,-   (c) Histamine receptor antagonists including H1 and H3 antagonists,-   (d) α₁- and α₂-adrenoceptor agonist vasoconstrictor sympathomimetic    agents for decongestant use,-   (e) muscarinic M3 receptor antagonists or anticholinergic agents,-   (f) PDE inhibitors, e.g. PDE3, PDE4 and PDE5 inhibitors,-   (g) Theophylline,-   (h) Sodium cromoglycate,-   (i) COX inhibitors both non-selective and selective COX-1 or COX-2    inhibitors (NSAIDs),-   (j) Oral and inhaled glucocorticosteroids, such as DAGR (dissociated    agonists of the corticoid receptor)-   (k) Monoclonal antibodies active against endogenous inflammatory    entities,-   (l) Anti-tumor necrosis factor (anti-TNF-α) agents,-   (m) Adhesion molecule inhibitors including VLA-4 antagonists,-   (n) Kinin-B₁- and B₂-receptor antagonists,-   (o) Immunosuppressive agents,-   (p) Inhibitors of matrix metalloproteases (MMPs),-   (q) Tachykinin NK₁, NK₂ and NK₃ receptor antagonists,-   (r) Elastase inhibitors,-   (s) Adenosine A2a receptor agonists,-   (t) Inhibitors of urokinase,-   (u) Compounds that act on dopamine receptors, e.g. D2 agonists,-   (v) Modulators of the NF_(κβ) pathway, e.g. IKK inhibitors,-   (w) modulators of cytokine signalling pathways such as p38 MAP    kinase, syk kinase or JAK kinase inhibitor-   (x) Agents that can be classed as mucolytics or anti-tussive, and-   (y) Antibiotics.

According to the present invention, combination of the compounds offormula (1) with:

-   -   H3 antagonists,    -   Muscarinic M3 receptor antagonists,    -   PDE4 inhibitors,    -   glucocorticosteroids,    -   Adenosine A2a receptor agonists,    -   Modulators of cytokine signalling pathyways such as p38 MAP        kinase or syk kinase, or,    -   Leukotriene antagonists (LTRAs) including antagonists of LTB₄,        LTC₄, LTD₄, and LTE₄,        are further preferred.

According to the present invention, combination of the compounds offormula (1) with:

-   -   glucocorticosteroids, in particular inhaled glucocorticosteroids        with reduced systemic side effects, including prednisone,        prednisolone, flunisolide, triamcinolone acetonide,        beclomethasone dipropionate, budesonide, fluticasone propionate,        ciclesonide, and mometasone furoate, or    -   muscarinic M3 receptor antagonists or anticholinergic agents        including in particular ipratropium salts, namely bromide,        tiotropium salts, namely bromide, oxitropium salts, namely        bromide, perenzepine, and telenzepine,        are further preferred.

It is to be appreciated that all references herein to treatment includecurative, palliative and prophylactic treatment. The description, whichfollows, concerns the therapeutic applications to which the compounds offormula (1) may be put.

The compounds of formula (1) have the ability to interact with the β2receptor and thereby have a wide range of therapeutic applications, asdescribed further below, because of the essential role which the β2receptor plays in the physiology of all mammals.

Therefore, a further aspect of the present invention relates to thecompounds of formula (1), or pharmaceutically acceptable salts, derivedforms or compositions thereof, for use in the treatment of diseases,disorders, and conditions in which the β2 receptor is involved. Morespecifically, the present invention also concerns the compounds offormula (1), or pharmaceutically acceptable salts, derived forms orcompositions thereof, for use in the treatment of diseases, disorders,and conditions selected from the group consisting of:

-   -   asthma of whatever type, etiology, or pathogenesis, in        particular asthma that is a member selected from the group        consisting of atopic asthma, non-atopic asthma, allergic asthma,        atopic bronchial IgE-mediated asthma, bronchial asthma,        essential asthma, true asthma, intrinsic asthma caused by        pathophysiologic disturbances, extrinsic asthma caused by        environmental factors, essential asthma of unknown or inapparent        cause, non-atopic asthma, bronchitic asthma, emphysematous        asthma, exercise-induced asthma, allergen induced asthma, cold        air induced asthma, occupational asthma, infective asthma caused        by bacterial, fungal, protozoal, or viral infection,        non-allergic asthma, incipient asthma, wheezy infant syndrome        and bronchiolytis,    -   chronic or acute bronchoconstriction, chronic bronchitis, small        airways obstruction, and emphysema,    -   obstructive or inflammatory airways diseases of whatever type,        etiology, or pathogenesis, in particular an obstructive or        inflammatory airways disease that is a member selected from the        group consisting of chronic eosinophilic pneumonia, chronic        obstructive pulmonary disease (COPD), COPD that includes chronic        bronchitis, pulmonary emphysema or dyspnea associated or not        associated with COPD, COPD that is characterized by        irreversible, progressive airways obstruction, adult respiratory        distress syndrome (ARDS), exacerbation of airways        hyper-reactivity consequent to other drug therapy and airways        disease that is associated with pulmonary hypertension,    -   bronchitis of whatever type, etiology, or pathogenesis, in        particular bronchitis that is a member selected from the group        consisting of acute bronchitis, acute laryngotracheal        bronchitis, arachidic bronchitis, catarrhal bronchitis, croupus        bronchitis, dry bronchitis, infectious asthmatic bronchitis,        productive bronchitis, staphylococcus or streptococcal        bronchitis and vesicular bronchitis,    -   acute lung injury,    -   bronchiectasis of whatever type, etiology, or pathogenesis, in        particular bronchiectasis that is a member selected from the        group consisting of cylindric bronchiectasis, sacculated        bronchiectasis, fusiform bronchiectasis, capillary        bronchiectasis, cystic bronchiectasis, dry bronchiectasis and        follicular bronchiectasis.

A still further aspect of the present invention also relates to the useof the compounds of formula (1), or pharmaceutically acceptable salts,derived forms or compositions thereof, for the manufacture of a drughaving a β2 agonist activity. In particular, the present inventionsconcerns the use of the compounds of formula (1), or pharmaceuticallyacceptable salts, derived forms or compositions thereof, for themanufacture of a drug for the treatment of β2-mediated diseases and/orconditions, in particular the diseases and/or conditions listed above.

As a consequence, the present invention provides a particularlyinteresting method to treat a mammal, including a human being, with aneffective amount of a compound of formula (1), or a pharmaceuticallyacceptable salt, derived form or composition thereof. More precisely,the present invention provides a particularly interesting method for thetreatment of a β2-mediated diseases and/or conditions in a mammal,including a human being, in particular the diseases and/or conditionslisted above, comprising administering said mammal with an effectiveamount of a compound of formula (1), its pharmaceutically acceptablesalts and/or derived forms.

The following examples illustrate the preparation of the compounds ofthe formula (1):

Preparation 1: Diethyl 2,2′-(1,3-phenylene)dlacetate

Acetyl chloride (12.5 ml, 175 mmol) was added to a suspension of2,2′-(1,3-phenylene)diacetic acid (50.0 g, 260 mmol) in ethanol (500 ml)and the resulting solution heated to reflux for 16 hours. The reactionwas cooled to room temperature and the solvent removed in vacuo. Theresidue was partitioned between saturated aqueous sodiumhydrogencarbonate (300 ml) and ethyl acetate (500 ml). The organic phasewas washed with water (200 ml), sat. aq. sodium chloride (300 ml), dried(sodium sulfate) and the solvent removed in vacuo to give the titlecompound as a pale yellow oil (63.5 g).

¹HNMR (CDCl₃ 400 MHz) δ: 1.31 (t, 6H), 3.65 (s, 4H), 4.20 (q, 4H),7.24-7.36 (m, 4H) ppm.

MS (electrospray): m/z 251 [M+H]⁺

Preparation 2: [3-(2-Oxo-propyl)-phenyl]-acetic acid ethyl ester

A solution of the diester from preparation 1 (44.3 g, 177 mmol) and2,2′-(1,3-phenylene)diacetic acid (59.2, 308 mmol) in ethanol (24 ml)and dioxan (290 ml) was treated dropwise with 12M hydrochloric acid (4.9ml, 58.8 mmol). The reaction mixture was stirred at reflux for 18 hoursbefore being allowed to cool and concentrated to low volume. Thereaction mixture was diluted with toluene (125 ml) and the resultingslurry filtered. The filtrate was concentrated in vacuo and the residuetaken up in water and basified with sodium bicarbonate until pH neutral.The mixture was diluted with ethyl acetate (200 ml) and the organiclayer was separated and washed with sodium bicarbonate solution (5×30ml) and saturated aqueous sodium chloride (50 ml). The combined aqueousextracts were acidified to pH 3 with 6M hydrochloric acid and extractedwith ether (3×30 ml). The organics were combined, dried (magnesiumsulphate) and concentrated in vacuo. The residue was triturated withpentane giving the title compound as a colourless solid 10.8 g.

¹HNMR (CD₃OD, 400 MHz) δ: 1.25 (t, 3H), 3.60 (m, 2H), 3.63 (m, 2H), 4.15(q, 2H), 7.18-7.32 (m, 4H) ppm.

MS (electrospray): m/z 245 [MNa]⁺

Preparation 3: [3-(2-Hydroxy-2-methyl-propyl)-phenyl]-acetic acid

Methyl magnesium chloride (51 ml of a 3M solution in tetrahydrofuran,153 mmol) was added dropwise to a stirred solution of the preparation 2(11.6 g, 51 mmol) (International Journal of Peptide and ProteinResearch, 1987, 29(3), 331) in tetrahydrofuran (300 ml) at 0° C. undernitrogen. The reaction was allowed to warm to room temperature overnightwith the formation of a thick white precipitate and then water (50 ml)and 2N hydrochloric acid (80 ml) were cautiously added. The aqueous wasextracted with ethyl acetate (2×300 ml) and the combined organics washedwith brine (50 ml), dried (sodium sulfate), and the solvent removed invacuo to furnish the title compound as a golden oil (11.2 g).

¹HNMR (CDC₃. 400 MHz) δ: 1.22 (6H, s), 2.75 (2H, s), 3.63 (2H, s),7.12-7.30 (4H, m).

MS (ESI): m/z 209 [M+H]⁺

Preparation 4:{3-[2-(2-Chloro-acetylamino)-2-methyl-propyl]-phenyl}-acetic acid

2-Chloroacetonitrile (8.8 ml, 140 mmol) was added to a solution of thealcohol from preparation 3 (16.0 g, 70 mmol), in acetic acid (33 ml).The resulting solution was cooled to 0° C., treated with concentratedsulphuric add (33 ml), and the reaction mixture allowed to warmgradually to room temperature. After 4 hours the reaction mixture waspoured onto ice and basified with solid sodium carbonate. The solutionwas extracted with ethyl acetate (2×500 ml) and the combined organicextracts dried (magnesium sulphate) and concentrated in vacuo to givethe title product as a colourless solid (19.0 g).

¹HNMR (CDCl₃, 400 MHz) δ: 1.36 (s, 6H), 3.02 (s, 2H), 3.62 (s, 2H), 3.95(s, 2H), 6.19 (m, 1H), 7.06-7.31 (m, 4H) ppm.

MS (electrospray): m/z 282 [M−H]⁻

Preparation 5: [3-(2-Amino-2-methyl-propyl)-phenyl]-acetic acid methylester

A solution of the amide from preparation 4 (5.1 g, 18 mmol), thiourea(1.6 g, 21 mmol) and acetic acid (18 ml) in ethanol (80 ml) was heatedto reflux under a nitrogen atmosphere for 16 hours. The reaction mixturewas allowed to cool to room temperature and filtered. The filtrate wasconcentrated in vacuo, the residue dissolved in methanol (150 ml),saturated with hydrogen chloride gas and the resulting solution heatedto reflux for 16 hours. The mixture was concentrated in vacuo and theresidue partitioned between ethyl acetate (200 ml) and 5% aqueous sodiumcarbonate solution (200 ml). The organic phase was washed with brine(100 ml), dried (magnesium sulphate) and concentrated in vacuo. Theresidue was purified by strong cation exchange resin, eluting withmethanol and then a 2M solution of ammonia in methanol, to elute theproduct. The eluent was concentrated in vacuo giving the title compoundas a yellow oil, 2.68 g.

¹HNMR (CDCl₃, 400 MHz) δ: 1.14 (s, 6H), 2.68 (s, 2H), 3.62 (s, 2H), 3.69(s, 3H), 7.08-7.16 (m, 3H), 7.23-7.27 (m, 1H) ppm.

MS (electrospray): m/z 222 [M+H]⁺

Preparation 6:N-{2-(benzyloxy)-5-[(1R)-2-bromo-1-hydroxyethyl]phenyl}methanesulfonamide

A solution of (1R)-1-[3-amino-4-(benzyloxy)phenyl]-2-bromoethanol (Org.Process Research and Development, 1998, 2, 96)_(30.8 g, 95.6 mmol) indichloromethane (300 ml) was treated with pyridine (9.3 ml, 115 mmol).The resulting solution was cooled to 5° C. and a solution ofmethanesulfonyl chloride (7.8 ml, 100.7 mmol) in dichloromethane (10 ml)added dropwise. The mixture was stirred at 5° C. for a further 30minutes and then allowed to warm gradually to room temperature over aperiod of 16 hours. The reaction mixture was washed with 2N hydrochloricacid (110 ml) and the organic phase separated, dried (magnesium sulfate)and the solvent removed in vacuo to give an orange oil. The residue wascrystallized from hot toluene (100 ml) to give the title compound as apale pink solid (33.7 g).

¹HNMR (DMSOd₆, 400 MHz) δ: 2.93 (s, 3H), 3.52-3.66 (m, 2H), 4.74 (m,1H), 5.19 (s, 2H), 7.11 (d, 1H), 7.19-7.22 (m, 1H), 7.33-7.36 (m, 2H),7.40-7.43 (m, 2H), 7.56 (d, 2H), 8.95 (s, 1H) ppm.

MS (electrospray): m/z 398/400 [M−H]−

Preparation 7: N-[2-(benzyloxy)-5-((1R)-2-bromo-1-{[tert-butyl(dimethyl)silyl]oxy}ethyl)phenyl]methanesulfonamide

A solution of the bromide of preparation 6 (21.5 g, 53.7 mmol) inN,N-dimethylformamide (125 ml) was treated with imidazole (4.16 g, 75.2mmol) and tert-butyl(dimethyl)silylchloride (9.73 g, 64.5 mmol) and theresulting solution left to stir at room temperature for 16 hours. Thereaction mixture was diluted with ethyl acetate (200 ml) and washed withwater (2×100 ml). The aqueous phases were combined and extracted withethyl acetate (100 ml). The combined organic extracts were washed with2N hydrochloric acid (100 ml), dried (magnesium sulphate) and reduced invacuo. The residue was suspended in pentane:ethyl acetate (200 ml, 1:1by volume) and the solvent evaporated. The residue was triturated withfurther pentane:ethyl acetate (200 ml, 1:1 by volume) and the resultingsolid filtered off and dried in vacuo to give the title compound as acolourless solid (23.7 g).

¹HNMR (CDCl₃, 400 MHz) δ: −0.07 (s, 3H), 0.11 (s, 3H), 0.89 (s, 9H),2.91 (s, 3H0, 4.80-4.83 (m, 1H), 6.80 (bs, 1H), 6.98 (d, 1H), 7.12 (d,1H), 7.36-7.44 (m, 5H), 7.52-7.54 (m, 1H) ppm.

Alternative Process for the Preparation of Preparation 7:

A solution of the bromide of preparation 6 (10 g, 24.98 mmol) wasdissolved in DCM (20 ml, 2 ml/g) and then imidazole (4.58 g, 37.47 mmol,1.5 eq) was added followed by TBDMSiCl (5.27 g, 34.97 mmol, 1.4 eq). Thereaction mixture was heated to reflux for 1 hour and then allowed tocool to 30° C. The mixture was diluted with isopropyl acetate (80 ml, 8ml/g) and then quenched with 2M HCl (50, 5 ml/g) and stirred vigorouslyfor 10 minutes. The phases were separated and the organic phase waswashed with water (50 ml, 5 ml/g). The organic phase was then reduced involume under reduced pressure at 45° C. to 25-30 ml. The solution wasthen cooled to room temperature and a suspension quickly formed and wasstirred at room temperature for 30 minutes. Heptane (20 ml, 2 ml/g) wasthen added over 10 minutes and the suspension was cooled to 5-10° C. andstirred for 1 hour. The suspension was then filtered and washed on thefilter paper with heptane (2×10 ml). The resulting filter cake was driedin a vacuum oven at 50° C. for 12 hours to give the title compound as awhite solid (11.05 g, 86% yield).

¹HNMR (CDCl₃, 400 MHz) δ: −0.07 (s, 3H), 0.11 (s, 3H), 0.89 (s. 9H),2.91 (s, 3H0, 4.80-4.83 (m, 1H), 6.80 (bs, 1H). 6.98 (d, 1H), 7.12 (d,1H), 7.36-7.44 (m, 5H), 7.52-7.54 (m, 1H) ppm.

Preparation 8: Methyl(3-{2-[((2R)-2-{4-(benzyloxy)-3-[(methylsulfonyl)amino]phenyl}-2-hydroxyethyl)amino]-2-methylpropyl}phenyl)acetate

The bromide of preparation 7 (36.0 g, 70.8 mmol) and the amine ofpreparation 5 (36.0 g, 153 mmol) were heated at 85° C. for 72 hours. Thereaction mixture was cooled to room temperature and purified by columnchromatography on silica gel eluting with pentane:ethyl acetate (50:50by volume) to yield the title product as a pale yellow oil (37.2 g).

¹HNMR (CDCl₃, 400 MHz) δ: −0.15 (s, 3H), 0.00 (s, 3H), 0.83 (s, 9H),1.01 (s, 3H), 1.04 (s, 3H), 2.57-2.97 (m, 7H), 3.59 (s, 2H), 3.68 (s,3H), 4.68-4.72 (m, 1H), 5.09 (s, 2H), 6.79 (bs, 1H), 6.95 (d, 1H),7.04-7.21 (m, 7H), 7.37-7.44 (m, 5H), 7.56 (d, 1H) ppm.

MS (APCI): m/z 655 [M+H]+

Preparation 9: methyl(3-{2-[((2R)-2-{[tert-butyl(dimethyl)silyl]oxy}-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetate

A solution of the benzyl protected alcohol of preparation 8 (36.8 g, 56mmol) in ethanol (550 ml) was treated with ammonium formate (16.0 g, 254mmol) and 20% palladium hydroxide on carbon (1.5 g). The resultingsuspension heated to 85° C. for 2 hours. After 2 hours further 20%palladium hydroxide on carbon (1.0 g) was added and heating continuedfor 1 hour. The reaction mixture was cooled to room temperature,filtered and the solvent removed in vacuo. The residue was partitionedbetween ethyl acetate (500 ml) and 2N aqueous ammonia (100 ml). Theorganic phase was separated, dried (magnesium sulfate) and the solventremoved in vacuo. The residue was purified by column chromatography onsilica gel eluting with dichloromethane:methanol:0.88:ammonia (95:5:0.5by volume) to yield the title product as a pale yellow oil (20.6 g).

¹HNMR (400 MHz, CDCl₃) δ: −0.17 (s, 3H), −0.05 (s, 3H), 0.80 (s, 9H),1.07 (s, 3H), 1.09 (s, 3H), 2.66-2.91 (m, 7H), 3.62 (d, 2H), 3.69 (s,3H), 4.71-4.74 (m, 1H), 6.58 (d, 1H), 6.88 (dd, 1H), 7.05-7.14 (m, 3H),7.21-7.25 (m, 1H), 7.30 (s, 1H) ppm.

MS (electrospray): m/z 565 [M+H]⁺

Preparation 10:(3-{2-[(2R)-2-(tert-Butyl-dimethyl-silanyloxy)-2-(4-hydroxy-3-methanesulfonylamino-phenyl)-thylamino]-2-methyl-propyl}-phenyl)-aceticacid

The ester of preparation 9 (20.6, 36 mmol) was dissolved intetrahydrofuran (150 ml) and the solution treated dropwise with 1Maqueous lithium hydroxide (72 ml, 72 mmol). The reaction mixture wasstirred at room temperature for 72 hours. The reaction mixture wasneutralised by addition of 1M hydrochloric acid (72 ml, 72 mmol) andconcentrated to low volume. The aqueous phase was decanted and theresidue washed with water (2×50 ml). The residue was redicolved intetrahydrofuran (50 ml) and toluene (50 ml) and the solvent removed invacuo to give the title compound as a pale brown foam (20.179).

¹HNMR (400 MHz, CD₃OD): −0.14 (s, 3H), 0.07 (s, 3H), 0.83 (s, 9H), 1.32(m, 6H), 2.93 (m, 5H), 3.23 (m, 2H), 3.54 (m, 2H), 4.94 (m, 1H), 6.91(d, 1H), 7.03-7.16 (m, 3H), 7.26 (m, 2H), 7.60 (m, 1H) ppm.

MS (electrospray): m/z 236 [M+H]⁺

Preparation 5a: [3-(2-amino-2-methyl-propyl)-phenyl]-acetic acid ethylester

A mixture of the amide from preparation 4 (151.4 g, 534 mmol), thiourea(48.7 g, 640 mmol) and acetic acid (303 ml) in ethanol (1.5 L) washeated to reflux under a nitrogen atmosphere for 5 hours. The reactionmixture was allowed to cool to room temperature and the suspensionconcentrated in vacuo. The residues were azeotroped with toluene (2×900mL) then treated with ethanol (1.5 L) and stirred for 1 hour. The solidprecipitate was removed by filtration, and the filtrate cooled in an icebath, treated with 98% sulphuric acid (227 mL) and stirred for 1 hour atambient temperature. The solution was concentrated in vacuo to removemost of the ethanol and adjusted to pH9 using aqueous sodiumbicarbonate. The solid precipitate was removed by filtration and washedwith water (300 mL) then ethyl acetate (1.0 L). The layers of thecombined biphasic filtrate and washes were separated and the aqueouslayer re-extracted with ethyl acetate (1.0 L+500 mL). The combined ethylacetate extracts were dried over magnesium sulphate, filtered andconcentrated in vacuo to give the title compound as a brown oil (89.5g).

¹H NMR (d₆-DMSO, 400 MHz) δ: 0.99 (s, 6H), 1.16 (t, 3H), 2.59 (s, 2H),3.61 (s, 2H), 4.06 (q, 2H), 7.06 (m, 3H), 7.21 (m, 1H)

Preparation 5b: [3-(2-amino-2-methyl-propyl)-phenyl]-acetic acid ethylester, dl-p-toluoyl-L-tartrate

A solution of the amine from preparation 5a (124.9 g, 531 mmol) inacetonitrile (1.0 L) was treated with a solution ofdi-p-toluoyl-L-tartaric acid (194.8 g, 504 mmol) in acetonitrile (750mL). The resulting slurry was stirred for 3 hours and the solidprecipitate isolated by filtration and washed with acetonitrile (2×250mL) to give the title compound as an off-white solid (210 g).

¹H NMR (d₆-DMSO, 400 MHz) δ: 1.13 (s, 6H), 1.17 (t, 3H), 2.34 (s, 6H),2.78 (s, 2H), 3.63 (s, 2H), 4.06 (q, 2H), 5.61 (s, 2H), 7.02 (d, 2H),7.15 (d. 1H), 7.25 (m, 5H), 7.80 (d, 4H)

Preparation 5c: [3-(2-Amino-2-methyl-propyl)-phenyl]-acetic acid ethylester

A solution of potassium carbonate (37.90 g, 274.22 mmol) in water (213ml) was added to a suspension of preparation 5b (42.62 g, 68.56 mmol) inpropionitrile (213 ml) and stirred until all solid had dissolved. Thephases were then separated and the propionitrile phase washed with water(107 ml). The solution was reduced in volume under reduced pressure toapproximately 30 ml to give the title compound as a propionitrilesolution. A sample was removed and concentrated to dryness to obtain aweight assay and yield was shown to be 81%.

¹H NMR (d₆-DMSO, 400 MHz) δ: 0.99 (s, 6H), 1.16 (t, 3H), 2.59 (s, 2H),3.61 (s, 2H), 4.06 (q, 2H), 7.06 (m, 3H), 7.21 (m, 1H)

Preparation 8a: Ethyl(3-{2-[((2R)-2-{[tert-butyl(dimethyl)silyl]oxy}-2-{4-benzyloxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetate

N-[2-(benzyloxy)-5-((1R)-2-bromo-1-{[tert-butyl(dimethyl)silyl]oxy}ethyl)phenyl]methanesulfonamide(14.34 g, 27.88 mmol) was added to the solution of preparation 5c (13.12g, 55.75 mmol) in propionitrile (15 ml). The mixture was then heated atreflux for 3 days. The solution was diluted with propionitrile (55 ml)and cooled to 20-25° C. The solution was washed with 1 M HCl_((aq)) (70ml) then water (35 ml) and the solution carried directly into the nextstep assuming 100% yield.

Preparation 9a: Ethyl(R)-2-(3-{2-[2-hydroxy-2-(4-benzyloxy-3-methanesulfonamidophenyl)ethylamino]-2-methylpropyl}phenyl)acetate

Triethylamine trihydrofluoride (9.1 ml, 8.99 g, 55.76 mmol) was added tothe solution of preparation 8a (18.64 g, 27.88 mmol) in propionitrile(72 ml). The solution was stirred at 20-25° C. for 3 hours. The solutionwas then quenched with 5M NH_(3(aq)) (72 ml) stirred for 10 minutes andthe phases separated. The propionitrile solution was then washed withwater (72 ml) and the solution carried directly into the next stepassuming 100% yield.

Preparation 10a:(R)-2-(3-{2-[2-hydroxy-2-(4-benzyloxy-3-methanesulfonamidophenyl)ethylamino]-2-methylpropyl}phenyl)aceticacid

A solution of sodium hydroxide (6.69 g, 167.28 mmol) in water (72 ml)was added to the solution of preparation 9a (15.47 g, 27.88 mmol) inpropionitrile (72 ml). The two phase mixture was then stirred vigorouslyfor 3 hours. The phases were allowed to separate and the aqueous phasewashed with fresh propionitrile (72 ml), then diluted with 1,4-dioxane(72 ml). The pH of the solution was then adjusted to pH 6-7 by theaddition of 37% w/w HCl_((aq)) and the resulting suspension was stirredfor one hour. The suspension was then filtered and washed on the filterpaper with water then dried giving the title compound as an off whitesolid (13.55 g, 92% over 3 steps).

¹HNMR (400 MHz, CD₃OD) δ: 1.33 (s, 3H), 1.35 (s, 3H), 2.89 (s, 3H), 2.96(s, 2H), 3.06-3.19 (m, 2H), 3.50 (s, 2H), 4.50 (m, 1H). 5.22 (s, 2H),7.08 (d, 1H), 7.13 (d, 1H), 7.19 (s, 1H), 7.24 (t, 2H), 7.27 (d, 1H),7.31 (d, 1H), 7.38 (t, 2H), 7.48 (d, 2H), 7.49 (s, 1H) ppm.

Preparation 10b:(R)-2-(3-{2-[2-hydroxy-2-(4-hydroxy-3-methanesulfonamidophenyl)ethylamino]-2-methylpropyl}phenyl)aceticacid sodium salt

A solution of sodium hydroxide (1.40 g, 35.05 mmol) in water (100 ml)was added to a suspension of preparation 10a (18.46 g, 35.05 mmol) inmethanol (600 ml). The mixture was hydrogenated over 20 wt % Palladiumhydroxide on carbon at 150 psi and 60° C. for 5 hours. The mixture wasfiltered to remove catalyst residues and then reduced in volume atreduced pressure to 100 ml. The mixture was distilled and replaced atreduced pressure to acetonitrile at constant volume. The resultingsuspension was filtered and washed on the paper with acetonitrile thendried to provide the title compound as an off white solid (15.34 g,95%).

¹HNMR (400 MHz, CD₃OD) δ: 1.07 (s, 3H), 1.09 (s, 3H), 2.70 (s, 2H),2.73-2.81 (m, 2H), 2.87 (s, 3H), 3.44 (s, 2H), 4.60-4.63 (m, 1H), 6.84(d, 1H), 6.92 (d, 1H), 7.04 (d, 1H), 7.11 (s, 1H), 7.14 (d, 1H), 7.15(t, 1H), 7.34 (s, 1H) ppm.

Said compounds of formula 10b can then be reacted with a suitable amineof formula NHR⁸-Q²-A (3)

in the presence of a conventional coupling agent such as1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride ordicyclohexylcarbodiimide in a suitable solvent such as pyridine,dimethylformamide or dimethylacetamide.

in order to obtain a compound of formula (1):

wherein R¹ and R² are methyl and n is 1.

Preparation 11: 1-(3-Bromophenyl)-2-methylpropan-2-ol

Methylmagnesium bromide (3M solution in diethylether, 51.6 ml, 155 mmol)was slowly added to a solution of 1-(3-bromo-phenyl)propan-2-one (15.0g, 70 mmol) in dry diethylether (200 ml) at 0° C. The resulting mixturewas left for 3 hours, then cooled to 0° C. and slowly quenched withsaturated aqueous ammonium chloride solution. The organic phase waswashed with brine, dried (sodium sulfate). The yellow oil was thenpurified by column chromatography on silica gel eluting withdichloromethane:pentane:methanol (90:5:5 by volume to afford a paleyellow oil (13.26 g).

¹H NMR (400 MHz, CDCl₃): δ 1.22 (s, 6H), 1.42 (bs, 1H), 2.74 (s, 2H),7.15 (m, 2H), 7.40 (m, 2H) ppm.

Preparation 12:N-[2-(3-Bromophenyl)-1,1-dimethylethyl]-2-chloroacetamide

Chloroacetonitrile (6.63 ml, 105 mmol) was added to a stirred solutionof the alcohol of preparation 11 (12.0 g, 52.0 mmol) in acetic acid (25ml) at room temperature. The resulting solution was cooled to 0° C. andconcentrated sulfuric acid (25 ml) was added keeping the temperature<10° C. The resulting solution was left to stir for 1 hour and thenpoured onto ice and basified by the addition of solid potassiumcarbonate. The product was extracted with ethyl acetate (2×500 ml), theorganics combined and washed with water (50 ml), dried (sodium sulfate)and the solvent removed in vacuo to afford the title compound as anorange solid (16.08 g).

¹H NMR (400 MHz, CDCl₃): δ 1.37 (s, 6H), 3.02 (s, 2H), 3.94 (s, 2H),6.17 (bs, 1H), 7.03-7.08 (d, 1H), 7.11-7.13 (t, 1H), 7.26 (s, 1H),7.32-7.39 (d, 1H) ppm.

LRMS (electrospray) m/z 306 [M+H]⁺

Preparation 13: [2-(3-Bromophenyl)-1,1-dimethylethyl]amine

A solution of the amide from preparation 12 (32.0 g, 105 mmol), thiourea(9.60 g, 126 mmol) and acetic acid (50 ml) in ethanol (250 ml) washeated to reflux overnight. The reaction mixture was cooled to roomtemperature and filtered, the filtrate was concentrated in vacuo andbasified using aqueous sodium hydroxide solution (1M, 450 ml). Theproduct was extracted with dichloromethane (2×500 ml) and the combinedorganics washed with brine (50 ml), dried (sodium sulfate) and thesolvent removed in vacuo to afford the title compound as a black oil (23g).

¹H NMR (400 MHz, CDCl₃): δ 1.12 (s, 6H), 1.84 (bs, 2H), 2.62 (s, 2H),7.08-7.16 (m, 2H), 7.32-7.36 (m, 2H)) pm.

LRMS (electrospray) m/z 228 [M+H]⁺

Preparation 14:N-[2-(Benzyloxy)-5-((1R)-2-{[2-(3-bromophenyl)-1,1-dimethylethyl]amino}-1-hydroxyethyl)phenyl]methanesulfonamide

The amine of preparation 13 (5.04 g, 22.3 mmol) was dissolved indichloromethane (20 ml) and treated withN-[2-(Benzyloxy)-5-((1S)-2-bromo-{[tert-butyl(dimethyl)silyl]oxy}ethyl)-phenyl]methanesulphonamide(WO 02/06258, pg. 36, example 14a) (11.90 g, 45.0 mmol). The resultingsolution heated at 90° C. until the solvent evaporated and then stirredat 90° C. for a further 16 hours. The reaction mixture was cooled toroom temperature and the residue was purified by column chromatographyon silica gel eluting with pentane:ethyl acetate (90:10) to yield thetitle product as a brown oil (8.36 g).

¹HNMR (CD₃OD, 400 MHz) δ: −0.14 (s, 3H), 0.04 (s, 3H), 0.84 (s, 9H),1.10 (s, 3H), 1.13 (s, 3H), 2.87 (s, 3H), 2.67-2.90 (m, 4H), 4.73-4.77(m, 1H), 5.25 (s, 2H), 7.12-7.23 (m, 4H), 7.36-7.48 (m, 6H), 7.53-7.55(m, 2H) ppm.

MS (electrospray): m/z 661/663 [M+H]⁺, 683/685 [M+H]⁺

Preparation 15: Methyl3-{2-[((2R)-2-{4-(benzyloxy)-3-[(methylsulfonyl)amino]phenyl}-2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)amino]-2-methylpropyl}benzoate

A solution of the bromide of preparation 14 (8.36 g, 12.6 mmol),(1,1′-bis(diphenylphosphino)ferrocene)dichloropalladium(II) (1.03 g,1.26 mmol) and triethylamine (3.5 ml, 25.1 mmol) in methanol was heatedat 100° C. under 100 psi carbon monoxide for 16 hours. The reactionmixture was cooled to room temperature, filtered and the solvent removedin vacuo. Purification by column chromatography on silica gel elutingwith dichloromethane:methanol: 0.880 ammonia (90:10:1) gave the titlecompound as an orange oil, 7.79 g (trace contamination with catalystresidues).

¹HNMR (400 MHz, CD₃OD) δ: −0.17 (s, 3H), 0.00 (s, 3H), 0.80 (s, 9H),1.12 (s, 3H), 1.15 (s, 3H), 2.67-2.92 (m, 3H), 3.96 (s, 3H), 4.73-4.77(m, 1H), 5.24 (s, 2H), 7.11 (d, 1H), 7.19 (dd, 1H), 7.36-7.48 (m, 6H),7.54 (d, 2H), 7.91-7.93 (m, 2H) ppm.

MS (electrospray) m/z 641 [M+H]⁺, 663 [M+Na]⁺

Preparation 16: Methyl3-{2-[((2R)-2-{[tert-butyl(dimethyl)silyl]oxy}-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}benzoate

Prepared from the ester of preparation 15 using the method ofpreparation 7 to give the title compound as a colourless oil.

¹HNMR (400 MHz, CD₃OD) δ: −0.21 (s, 3H), −0.05 (s, 3H), 0.75 (s, 9H),1.08 (s, 3H), 1.12 (s, 3H), 2.62-2.88 (m, 7H), 3.92 (s, 3H), 4.64-4.69(m, 1H), 6.84 (d, 1H), 7.03 (dd, 1H), 7.35-7.36 (m, 1H), 7.39-7.42 (m,2H), 7.87-7.89 (m, 2H) ppm.

MS (electrospray) m/z 551 [M+H]⁺, 573 [M+Na]⁺

Preparation 17:3-{2-[((2R)-2-{[tert-Butyl(dimethyl)silyl]oxy}-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}benzoicacid

Prepared from the ester of preparation 16 to using the method ofpreparation 8 to give the title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD) δ: −0.14 (s, 3H), 0.04 (s, 3H), 0.82 (s, 9H),1.23 (s, 3H), 1.24 (s, 3H), 2.88-2.96 (m, 5H), 3.00-3.14 (m, 2H),4.83-4.87 (m, 1H), 6.89 (d, 1H), 7.07 (dd, 1H), 7.24-7.26 (m, 1H), 7.32(t, 1H), 7.37 (s, 1H), 7.82 (s, 1H), 7.86 (d, 1H) ppm.

MS (electrospray) m/z 537 [M+H]⁺, 559 [M+Na]⁺

Preparation 18-53

A solution of the appropriate carboxylic acid (preparation 10 or 17)(0.15 mmol), 1-hydroxybenzotriazole hydrate (22 mg, 0.16 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (34 mg, 0.18mmol) and N-ethyldiisopropylamine (130 μL, 0.73 mmol) inN,N-dimethylformamide (2 ml) was treated with the appropriate amine(0.23 mmol) and the reaction mixture shaken at room temperature for 18hours. The reaction mixture was concentrated in vacuo and the residuepartitioned between dichloromethane (3 ml) and water (1 ml). The phaseswere separated and the organic layer washed with brine (1 ml), dried(sodium sulphate) and concentrated in vacuo. The residue was purified bycolumn chromatography on silica gel eluting withdichloromethane:methanol:0.88 ammonia (98:2:0 changing to 94:6:0.5 byvolume) to yield the desired product.

Alternatively, the following process can be used for the synthesis ofpreparations 18 to 53:

A solution of appropriate carboxylic acid from preparation 10 or 17(5.08 mmol) in N,N-dimethylformamide (60 ml) is treated with1-hydroxybenzotriazole hydrate (0.755 g, 5.59 mmol), the appropriateamine (5.08 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (1.07 g, 5.59 mmol) and triethlyamine (1.49 ml, 10.67mmol). The resulting suspension is left to stir at room temperature for18 hours. The solvent is removed in vacuo and the residue partitionedbetween dichloromethane (100 ml) and sat. aq. sodium bicarbonate (50ml). The organic phase is separated and the aqueous phase extracted withdichloromethane:methanol (95:5 by volume, 2×20 ml). The combined organicextracts are separated, washed with saturated aqueous sodium chloride(100 ml), dried (sodium sulfate) and the solvent removed in vacuo. Theresidue is purified by column chromatography on silica gel eluting withdichloromethane:methanol:0.88 ammonia (95:5:0.5 by volume) to yield thedesired compound.

Preparations 18 to 47

No Q¹ Data 18

¹HNMR (CDCl₃, 400 MHz) δ: −0.19 (s, 3 H), −0.11(s, 3 H), 0.75 (s, 9 H),1.03 (s, 3 H), 1.05(s, 3 H), 2.57-2.85 (m, 7 H), 3.57 (m, 2 H), 3.74(s,3 H), 4.31 (m, 2 H), 4.65 (m, 1 H). 6.22 (m,1 H), 6.63 (d, 1 H), 6.67(m, 1 H), 6.76 (m, 1 H),6.82 (d, 1 H), 7.03 (m, 2 H), 7.12 (m, 1 H),7.22(m, 1 H), 7.34 (m, 1 H)MS (electrospray) m/z 686 [M + H]⁺ 19

¹HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.02(s, 3 H), 0.79 (s, 9 H),1.02 (s, 3 H), 1.04(s, 3 H), 2.60-2.71 (m, 3 H), 2.82-2.87 (4 H, m),3.52(s, 2 H), 4.36 (s, 2 H), 4.65-4.68 (m, 1 H),6.81-6.85 (m, 3 H),6.99-7.03 (m, 2 H), 7.10 (s,1 H), 7.16-7.22 (m, 2 H), 7.24-7.26 (d, 2H),7.35 (d, 1 H), 7.40 (d, 2 H), 7.45 (d, 2 H).MS (electrospray) m/z 730[M − H]⁻, 732 [M + H]⁺ 20

¹HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.02(s, 3 H), 0.80 (s, 9 H),1.02 (s, 3 H), 1.05(s, 3 H), 2.61-2.71 (m, 3 H), 2.83-2.87 (4 H, m),3.52(s, 2 H), 4.29 (s, 2 H), 4.66-4.69 (m, 1 H),6.72 (d, 1 H), 6.84 (d, 1H), 7.00-7.05 (m, 4 H),7.08 (s, 1 H), 7.14-7.22 (m, 2 H), 7.35 (s, 1H)MS (APCl) m/z 688 [M-H]⁻, 690 [M + H]⁺ 21

¹HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.02(s, 3 H), 0.79 (s, 9 H),1.01 (s, 3 H), 1.04(s, 3 H), 2.14 (s, 6 H), 2.60-2.70 (m, 3 H),2.82-2.87(4 H, m) 3.48 (s, 2 H), 4.18 (s, 2 H), 4.65-4.68(m, 1 H), 6.77(s, 2 H), 6.83 (d, 1 H), 6.98-7.02(m, 2 H), 7.08 (s, 1 H), 7.14-7.21 (m,2 H),7.35 (s, 1 H).MS (APCl) m/z 684 [M + H]⁺ 22

¹HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.03(s, 3 H), 0.79 (s, 9 H),0.89 (s, 3 H), 0.91(s, 3 H), 2.50-2.60 (m, 3 H), 2.76-2.81 (m, 1 H),2.85(s, 3 H), 3.48 (s, 2 H), 4.64 (dd, 1 H), 4.78(s, 2 H), 6.85 (d, 1 H),6.95-6.97 (m, 2 H), 7.01(d, 1 H), 7.09-7.14 (m, 3 H), 7.26 (dd, 1 H),7.35(s, 1 H), 7.39 (dd, 1 H), 7.68 (d, 1 H), 7.72 (d,1 H), 7.89 (d, 1H).MS (electrospray) m/z 706 [M + H]⁺, 704 [M − H]⁻ 23

¹HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.02(s, 3 H), 0.79 (s, 9 H),1.00 (s, 3 H), 1.02(s, 3 H), 2.60-2.71 (m, 3 H), 2.82-2.86 (m, 1 H),2.86(s, 3 H), 3.54 (s, 2 H), 4.45 (s, 2 H), 4.68(dd, 1 H), 6.85 (d, 1 H),7.03 (m, 4 H), 7.09 (s,1 H), 7.17-7.27 (m, 3 H), 7.35 (s, 1 H),7.54-7.61(m, 3 H).MS (electrospray) m/z 704 [M − H]⁻ 24

¹HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.02(s, 3 H), 0.79 (s, 9 H),0.99 (s, 3 H), 1.02(s, 3 H), 2.56-2.66 (m, 3 H), 2.80-2.83 (m, 1 H),2.85(s, 3 H), 3.53 (s, 2 H), 4.40 (s, 2 H), 4.67(dd, 1 H), 6.81 (d, 2 H),6.84 (d, 1 H), 7.01 (d,2 H), 7.09 (s, 1 H), 7.13 (d, 1 H), 7.18-7.20(m,2 H), 7.29 (dd, 1 H), 7.33-7.40 (m, 5 H).MS (electrospray) m/z 730 [M− H]⁻, 732 [M + H]⁺,754 [M + Na]⁺ 25

¹HNMR (400 MHz, CD₃OD) δ: −0.18 (s, 3 H), −0.01(s, 3 H), 0.80 (s, 9 H),1.01 (s, 3 H), 1.04(s, 3 H), 2.58-2.68 (m, 2 H), 2.86 (s, 3 H), 3.34(s,2 H), 3.53 (s, 2 H), 4.41 (s, 2 H), 4.66-4.69(m, 1 H), 6.73 (dd, 1 H),6.84 (d, 1 H), 6.94-6.97(m, 2 H), 6.99-7.02 (m, 2 H), 7.08 (s, 1 H),7.16-7.21(m, 4 H), 7.31 (t, 1 H), 7.34 (d, 1 H),7.40-7.42 (m, 2 H)ppm.MS (electrospray) m/z 730 [M − H]^(−.) 26

¹HNMR (400 MHz, CD₃OD) δ: −0.17 (s, 3 H),0.00 (s, 3 H), 0.17 (s, 6 H),0.80 (s, 9 H), 0.98(s, 9 H), 1.09 (s, 3 H), 1.12 (s, 3 H), 1.21 (s,6 H),2.67-2.78 (m, 3 H), 2.87 (s, 3 H), 2.91-2.96(m, 1 H), 3.36 (s, 2 H),3.43 (s, 2 H), 4.71-4.75(m, 1 H), 6.73 (d, 2 H), 6.86 (d, 1 H),7.03-7.06(m, 4 H), 7.15-7.22 (m, 3 H), 7.37 (d, 1 H).MS (APCl) m/z 812[M + H]⁺ 27

¹HNMR (400 MHz, CD₃OD) δ: −0.18 (s, 3 H), −0.01(s, 3 H), 0.80 (s, 9 H),1.03 and 1.08 (2 s,3 H), 1.01 and 1.07 (2 s, 3 H), 1.25-1.29 (2 t,3 H),2.65-2.69 (m, 2 H), 2.87 and 2.88 (2 s,3 H), 2.92 (q, 2 H), 3.75 and3.82 (2 s, 2 H),4.04 (s, 2 H), 4.55 and 4.57 (2 s, 2 H), 4.68-4.71(m, 1H), 6.86 (dd, 1 H), 6.98 (d, 2 H), 7.03(dd, 1 H), 7.10 (d, 1 H),7.13-7.15 (m, 1 H), 7.20(d, 1 H), 7.24 (dd, 1 H), 7.36 (dd, 1 H).MS(electrospray) m/z 750/752 [M − H]⁻ 28

¹HNMR (400 MHz, CD₃OD) δ: −0.21 and −0.19(2 s, 3 H), −0.05 and −0.03 (2s, 3 H), 0.76 and0.78 (2 s, 9 H), 0.96 and 0.98 (2 s, 3 H), 0.93and 0.97(2 s, 3 H), 2.52-2.56 (m, 2 H), 2.60-2.64(m , 2 H), 2.86 (s, 3 H), 2.85and 3.02 (2 s,2 H), 3.34 (s, 3 H), 3.84 (s, 2 H), 6.84 (d, 2H),6.97-7.01 (m, 1 H), 7.02-7.03 (m, 1 H), 7.05 (d,1 H), 7.11 (d, 1 H),7.13-7.15 (m, 1 H), 7.17-7.20(m, 1 H), 7.27-7.31 (m, 1 H), 7.35-7.36(m,1 H), 7.56-7.60 (m, 2 H), 7.65-7.68 (m, 1 H),7.97 (d, 1 H).MS(electrospray) m/z 750/752 [M − H]⁻ 29

¹HNMR (400 MHz, CD₃OD) δ: −0.18 (s, 3 H), −0.01(s, 3 H), 0.80 (s, 9 H),1.02 (s, 3 H), 1.04(s, 3 H), 2.58-2.68 (m, 2 H), 2.86 (s, 3 H), 3.34(s,2 H), 3.52 (s, 2 H), 4.40 (s, 2 H), 4.66-4.69(m, 1 H), 6.84 (m, 3 H),6.97-7.02 (m, 2 H),7.07-7.18 (m, 6 H), 7.28 (t, 1 H), 7.34 (d, 1 H),7.38(s, 1 H), 7.40-7.42 (m, 1 H).MS (electrospray) m/z 730 [M − H]⁻ 30

¹HNMR (400 MHz, CD₃OD) δ: −0.18 (s, 3 H),0.00 (s, 3 H), 0.80 (s, 9 H),0.95-1.05 (m, 6 H),2.50-2.90 (m, 9 H), 3.68 and 3.74 (2 t, 2 H),3.82 (s.2 H), 4.57 and 4.59 (2 s, 2 H), 4.62-4.69(m, 1 H), 6.50-6.62 (m, 2 H),6.78-7.23(m, 7 H), 7.37-7.39 (m, 1 H).MS (APCl) m/z 680 [M − H]⁻ 31

¹HNMR (400 MHz, CD₃OD) δ: −0.18 (s, 3 H),0.01 (s, 3 H), 0.81 (s, 9 H),1.07 (s, 3 H), 1.10(s, 3 H), 1.45-1.62 (m, 6 H), 2.45-2.54 (m, 2H),2.62-2.76 (2 H, m), 2.86 (s, 3 H), 3.15-3.22 (m,2 H), 3.44 (s, 2 H),4.65-4.70 (m, 1 H), 6.62-6.65(m, 2 H), 6.83 (d, 1 H), 6.93 (d, 1H),6.96-7.22 (m, 6 H), 7.36 (s, 1 H).MS (electrospray) m/z 698 [M + H]⁺,696 [M − H]⁻ 32

¹HNMR (400 MHz, CD₃OD) δ: −0.94 (s, 3 H),0.00 (s, 3 H), 0.80 (s, 9 H),1.08 (s, 3 H), 1.12(s, 3 H), 2.62-2.77 (m, 6 H), 2.85 (s, 3 H), 3.36(t,2 H), 3.42 (s, 2 H), 4.64-4.70 (m, 1 H), 6.63(d, 2 H), 6.84 (d, 1 H),9.95 (d, 2 H), 7.00-7.06(3 H, m), 7.09 (d, 1 H), 7.69 (t, 1 H), 7.37(s,1 H).MS (electrospray) m/z 668 [M − H]⁻ 33

¹HNMR (400 MHz, CD₃OD) δ: −0.16 (s, 3 H),0.01 (s, 3 H), 0.82 (s, 9 H),1.05 (s, 3 H), 1.08(s, 3 H), 2.64-2.74 (m, 3 H), 2.85-2.90 (m, 4 H),3.52(s, 2 H), 4.36 (s, 2 H), 4.68-4.70 (m, 1 H),6.65 (d, 1 H), 6.80-6.81 (m,1 H), 6.86 (d, 1 H),7.04 (d, 2 H), 7.10-7.12 (m, 2 H), 7.16-7.23 (m,2H), 7.37-7.38 (m, 1 H).MS (electrospray) m/z 688 [M − H]⁻ 34

¹HNMR (400 MHz, CD₃OD) δ: −0.16 (s, 3 H),0.02 (s, 3 H), 0.82 (s, 9 H),1.08 (s, 3 H), 1.11(s, 3 H), 2.68-2.78 (m, 3 H), 2.89-2.96 (m, 4 H),3.52(s, 2 H), 4.21 (s, 2 H), 4.71-4.74 (m, 1 H),6.86 (d, 1 H), 7.03-7.07 (m,3 H), 7.11 (s, 2 H),7.15-7.24 (m, 2 H), 7.36-7.37 (m, 1 H).MS (APCl) m/z724/726 [M + H]⁺,722/724 [M − H]⁻ 35

¹HNMR (400 MHz, CD₃OD) δ: −0.16 (s, 3 H),0.01 (s, 3 H), 0.82 (s, 9 H),1.09 (s, 3 H), 1.11(s, 3 H), 2.68-2.78 (m, 3 H), 2.89-2.96 (m, 4 H),3.53(s, 2 H), 4.21 (s, 2 H), 4.71-4.74 (m, 1 H),6.79 (d, 1 H), 6.84-6.87 (m,2 H), 7.04-7.11 (m,3 H), 7.17-7.25 (m, 2 H), 7.39 (s, 1 H).MS (APCl) m/z724/726 [M + H]⁺,722/724 [M − H]⁻ 36

¹HNMR (400 MHz, CD₃OD) δ: −0.20 (s, 3 H), −0.04(s, 3 H), 0.78 (s, 9 H),0.93 (s, 3 H), 0.95(s, 3 H), 2.54-2.63 (m, 4 H), 2.84 (s, 3 H), 3.46(s,2 H), 4.63-4.67 (m, 3 H), 6.72 (d, 1 H), 6.84(d, 1 H), 6.95-7.02 (m, 2H), 7.10-7.14 (m, 2 H),7.20 (d, 1 H), 7.35-7.39 (m, 4 H), 7.82-7.85 (m,1H), 7.92 (s, 1 H), 8.20-8.22 (m, 1 H).MS (APCl) m/z 706 [M + H]⁺, 728[M + Na]⁺,704 [M − H]⁻ 37

^(‘)HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.02(s, 3 H), 0.80 (s, 9H), 1.03 (s, 3 H), 1.06(s, 3 H), 2.63-2.73 (m, 2 H), 2.86 (s, 3 H),3.34(s, 2 H), 3.53 (s, 2 H), 4.34 (s, 2 H), 4.66-4.69(m, 1 H), 6.84 (d,1 H), 6.87-6.90 (m, 2 H), 6.96(s, 1 H), 7.01-7.04 (m, 2 H), 7.09 (s, 1H),7.15-7.22 (m, 2 H). 7.35 (d, 1 H) ppm.MS (APCl) m/z 724 [M + H]⁺, 746[M + Na]⁺ 38

¹HNMR (400 MHz, CD₃OD) δ: −0.19 and −0.18(2 s, 3 H), −0.02 and −0.01 (2s, 3 H), 0.80 (s,9 H), 1.03-1.07 (m, 6 H), 1.09 (t, 3 H), 2.64-2.71(m, 2H), 2.87 (s, 3 H), 3.37 (q, 2 H), 3.61 and3.69 (2 s, 2 H), 3.80 and 3.82(2 s, 2 H), 4.50and 4.53 (2 s, 1 H), 4.67-4.71 (m, 1 H), 6.46-6.51(m, 1H), 6.62-6.67 (m, 1 H), 6.69-6.74(m, 1 H), 6.84 (dd, 1 H), 6.97-7.25 (m,5 H),7.35 (d, 1 H) ppm.MS (APCl) m/z 718/720 [M + H]⁺ 39

¹H NMR (400 MHz, CDCl₃) δ: (rotamers) −0.21and −0.18 (2 s, 3 H), −0.02and −0.01 (2 s,3 H), 0.76 (s, 9 H), 1.24 (s, 6 H), 2.88 (m, 3 H),2.94and 2.95 (2 s, 3 H), 2.98 (m, 4 H), 3.69and 3.76 (2 s, 2 H), 4.53 (m, 2H), 4.62-4.70 (m,1 H), 6.60-7.23 (m, 10 H).LRMS (electrospray): m/z [M +H]⁺704, [M + Na]⁺ 726. 40

¹H NMR (400 MHz, CD₃OD) δ: (rotamers) −0.18and −0.19 (2 s, 3 H), −0.03and −0.02 (2 s,3 H), 0.79 (s, 9 H), 1.00-1.05 (m, 6 H), 2.61-2.66(m, 2H), 2.87 (s, 3 H), 2.92 and 2.97 (2 s,3 H), 3.77 and 3.82 (2 s, 2 H),4.58 and 4.65(2 s, 2 H), 4.66-4.69 (m, 1 H), 4.80 (s, 2 H), 6.71(d, 1H), 6.84 (d, 1 H), 6.91 (s, 2 H), 6.96-7.03(m, 2 H), 7.05 (s, 1 H), 7.12(d, 1 H), 7.19-7.24(m, 1 H), 7.35 (s, 1 H) ppm.LRMS (electrospray): m/z738 [M − H]⁺ 41

¹H NMR (400 MHz, CD₃OD): δ −0.13 (s, 3 H),0.04 (s, 3 H), 0.85 (s, 9 H),1.10 (s, 3 H), 1.12(s, 3 H), 2.69-2.78 (m, 3 H), 2.89-2.94 (m, 4 H),3.50(s, 2 H), 4.28-4.29 (m, 2 H), 4.71-4.74 (m,1 H), 6.87-6.92 (m, 3 H),7.04-7.09 (m, 3 H),7.10 (s, 1 H), 7.13-7.26 (m, 4 H), 7.27-7.32 (m,3 H),7.39 (d, 1 H) ppm.LRMS (electrospray): m/z [M + H]⁺ 733, [M − H]⁻731. 42

¹H NMR (400 MHz, CD₃OD): δ −0.12 (s, 3 H),0.05 (s, 3 H), 0.85 (s, 9 H),1.15 (s, 3 H), 1.17(s, 3 H), 2.74-2.84 (m, 3 H), 2.92 (s, 3 H),2.95-3.02(m, 1 H), 3.53 (s, 2 H), 4.34 (s, 2 H), 4.75-4.79(m, 1 H),6.74-6.81 (m, 3 H), 6.90 (d, 1 H),7.06-7.10 (m, 2 H), 7.14 (s, 1 H),7.16-7.32 (m,7 H), 7.40 (d, 1 H) ppm.LRMS (APCl): m/z [M + H]⁺ 733, [M −H]⁻ 731. 43

¹H NMR (400 MHz, CD₃OD): δ −0.19 (s, 3 H), −0.02(s, 3 H), 0.79 (s, 9 H),1.01 (s, 3 H), 1.04(s, 3 H), 2.14 (s, 6 H), 2.60-2.70 (m, 3 H), 2.85(s,3 H), 2.82-2.87 (m, 1 H), 3.48 (s, 2 H), 4.18(s, 2 H), 4.65-4.68 (m, 1H), 6.77 (s, 2 H), 6.83(d, 1 H), 6.98-7.02 (m, 2 H), 7.08 (s, 1H),7.14-7.21 (m, 2 H), 7.35 (d, 1 H) ppm.LRMS (APCl): m/z [M + H]⁺ 68444

¹HNMR (400 MHz, CDCl₃) δ: −0.81 (s, 3 H),0.00 (d, 3 H), 0.81 (s, 9 H),0.95-1.05 (m, 6 H),2.50-2.90 (m, 9 H), 3.60-3.82 (m, 4 H), 4.48-4.70(m,3 H), 6.40-6.60 (m, 2 H), 6.80-7.41(m, 8 H) ppm.MS (electrospray) m/z682 [M + H]⁺, 704[M + Na]⁺ 45

¹HNMR (400 MHz, DMSO_(d6)) δ: −0.90 (m, 3 H),0.05 (m, 3 H), 0.8 (m, 9H), 1.21-1.29 (m, 6 H),2.60-2.75 (m, 2 H), 2.92 (s, 3 H), 2.98 (m, 2H),3.30 (m, 2 H), 3.71-4.00 (m, 4 H), 4.61-4.81(m, 2 H), 4.89-4.91 (m, 1H), 6.51-6.62 (m, 2 H),6.92-7.49 (m, 8 H), 7.72-7.80 (m, 1 H) ppm.MS(electrospray) m/z 60 [M − H]⁻ 46

¹HNMR (400 MHz, CDCl₃) δ: −0.16 (s, 3 H), −0.05(s, 3 H), 0.80 (s, 9 H),1.10 (s, 3 H), 1.18(s, 3 H), 2.63 (d, 1 H), 2.78 (m, 2 H), 2.83 (m,4 H),3.60 (d, 1 H), 3.62 (d, 1 H), 3.80 (m, 2 H),4.70 (m, 1 H), 5.08 (m, 1H), 6.60 (m, 2 H), 6.81(d, 1 H), 7.08 (m, 2 H), 7.20 (m, 7 H) ppm.MS(APCi) m/z 668 [M − H]⁻ 47

¹HNMR (400 MHz, CDCl₃) δ: −0.18 (s, 3 H), −0.10(s, 3 H), 0.78 (s, 9 H),1.10 (s, 3 H), 1.18(s, 3 H), 2.63 (d, 1 H), 2.80 (m, 6 H), 3.60 (d,1 H),3.78 (m, 3 H), 4.75 (m, 1 H), 5.10 (m, 1 H),6.38 (d, 1 H), 5.78 (d, 2H), 7.05 (m, 1 H), 7.20(m, 8 H) ppm.MS (APCi) m/z 670 [M + H]⁺

Preparations 48-53

48

¹HNMR (400 MHz, CD₃OD) δ: −0.16 (s, 3 H),0.00 (s, 3 H), 0.82 (s, 9 H),1.13 (s, 3 H), 1.17(s, 3 H), 2.68-2.93 (m, 7 H), 4.64 (s, 2 H),4.69-4.74(m, 1 H), 6.80-6.87 (m, 2 H), 7.05 (dd,1 H), 7.30-7.47 (m, 7 H), 7.54(d, 2 H),7.75-7.78 (m, 3 H) ppm.MS (electrospray) m/z 718 [M + H]⁺ 49

¹HNMR (400 MHz, CD₃OD) δ: −0.15 (s, 3 H), −0.01(s, 3 H), 0.22 (s, 6 H),0.83 (s, 9 H), 1.02(s, 9 H), 1.11 (s, 3 H), 1.13 (s, 3 H 0, 1.41 (s,6H), 2.67-2.92 (m, 7 H), 3.59 (s, 2 H),4.70-4.73 (m, 1 H), 6.83-6.90 (m,3 H), 7.07 (dd,1 H), 7.32-7.40 (m, 5 H), 7.55-7.58 (m, 2 H)ppm.MS(electrospray) m/z 799 [M + H]⁺ 50

¹HNMR (400 MHz, CD₃OD) δ: −0.19 (s, 3 H), −0.03(s, 3 H), 0.79 (s, 9 H),2.67-2.92 (m, 7 H),4.68-4.71 (m, 3 H), 6.86-6.88 (m, 3 H), 7.04 (d,1 H),7.31-7.33 (m, 1 H), 7.37-7.49 (m, 7 H),7.61 (m, 1 H), 7.76-7.80 (m, 2 H)ppm.MS (electrospray) m/z 718 [M + H]⁺, 740[M + H]⁺ 51

¹HNMR (400 MHz, CD₃OD) δ: −0.27 (s, 3 H), −0.10(s, 3 H), 0.71 (s, 9 H),0.99 (s, 3 H), 1.02(s, 3 H), 2.02 (s, 3 H), 2.17 (s, 3 H), 2.56-2.81(m,9 H), 3.39-3.43 (m, 2 H), 4.59-4.63 (m, 1 H),6.48 (s, 1 H), 6.77-6.79(m, 2 H), 6.95-6.97 (m,1 H), 7.22-7.29 (m, 3 H), 7.55-7.59 (m, 2H)ppm.MS (electrospray) m/z 685 [M + H]⁺ 52

¹HNMR (400 MHz, CD₃OD) δ: −0.18 (s, 3 H), −0.01(s, 3 H), 0.80 (s, 9 H),1.09 (s, 3 H), 1.12(s, 3 H), 2.14 (s, 3 H), 2.28 (s, 3 H), 2.65-2.92(m,9 H), 3.47-3.51 (m, 2 H), 4.68-4.71 (m, 1 H),6.55 (d, 1 H), 6.83-6.87(m, 2 H), 7.03-7.06 (m,1 H), 7.32-7.38 (m, 3 H), 7.64-7.68 (m, 2H)ppm.MS (electrospray) m/z 685 [M + H]⁺ 53

¹HNMR (400 MHz, CD₃OD) δ: −0.27 (s, 3 H), −0.07(s, 3 H), 0.75 (s, 9 H),1.03 (s, 3 H), 1.06(s, 3 H), 2.11 (s, 3 H), 2.60-2.85 (m, 9 H),3.47-3.51(m, 2 H), 4.63-4.66 (m, 1 H), 6.62 (d, 1 H),6.80-6.86 (m, 2 H),6.92 (m, 1 H), 6.98-7.01(m, 1 H), 7.26-7.33 (m, 3 H), 7.57-7.61 (m, 2H)ppm.MS (electrospray) m/z 671 [M + H]⁺

Preparation 54:N-(4-bromobenzyl)-2-(3-{2-[((2R)-2-{[tert-butyl(dimethyl)silyl]oxy}-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide

Prepared using the procedure for preparation 18 using the acid frompreparation 10 and (4-bromobenzyl)amine to give the title compound as ayellow gum.

¹HNMR (400 MHz, CDCl₃): δ −0.18 (s, 3H), 0.00 (s, 3H), 0.81 (s, 9H),1.02 (s, 3H), 1.04 (s, 3H), 2.61-2.72 (m. 4H), 2.83 (s, 3H), 3.53 (s,2H), 4.33 (s, 2H), 4.65-4.70 (m, 1H), 6.83-6.86 (d, 1H), 7.00-7.44 (m,10H) ppm.

MS (electrospray) m/z 720 [M+H]⁺, 742 [M+H]⁺

Preparation 55:2-(3-{2-[((2R)-2-{[tert-butyl(dimethyl)silyl]oxy}-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(3′-hydroxybiphenyl-4-yl)methyl]acetamide

A solution of the bromide from preparation 56 (0.50 g, 0.70 mmol),(3-hydroxyphenyl)boronic acid (0.19 g, 1.4 mmol),(1,1′-bis(diphenylphosphino) ferrocene)dichloropalladium(II) (36 mg,0.04 mmol) in N,N-dimethylformamide (8 ml) was treated with 2M aqueoussodium carbonate (2 ml) and the resulting suspension heated to 80° C.for 16 hours. The reaction mixture was cooled to room temperature andthe solvent removed in vacuo. The residue was azeotroped with toluene(50 ml), redisolved in ethyl acetate (50 ml) and neutralised with 1Naqueous hydrochloric acid (to pH7). The organic layer was separated andthe aqueous extracted with further ethyl acetate (2×50 ml). The combinedorganic extracts were washed with water (100 ml), saturated aqueoussodium chloride (100 ml), dried (sodium sulfate) and the solvent removedin vacuo to give an orange gum (515 mg) which was used without furtherpurification.

¹HNMR (400 MHz, CD₃OD): δ −0.13 (s, 3H), 0.04 (s, 3H), 0.84 (s, 9H),1.11 (s, 3H), 1.13 (s, 3H), 2.74-2.97 (m, 7H), 3.55-3.63 (m, 2H),4.42-4.45 (m, 2H), 4.73-4.76 (m, 1H), 6.89-6.94 (m, 3H), 7.15-7.30 (m,9H), 7.41 (d, 1H), 7.51 (s, 1H), 7.53 (s, 1H) ppm.

MS (electrospray) m/z 732 [M+H]⁺, 754 [M+H]⁺

Preparation 56:2-(3-{2-[((2R)-2-{[tert-butyl(dimethyl)silyl]oxy}-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(2′-hydroxybiphenyl-4-yl)methyl]acetamide

Prepared from (2-hydroxyphenyl)boronic acid using the method ofpreparation 55 to give the title compound as a brown oil.

¹HNMR (400 MHz, CD₃OD): δ −0.13 (s, 3H), 0.04 (s, 3H), 0.84 (s, 9H),1.12 (s, 3H), 1.14 (s, 3H), 2.72-2.99 (m, 7H), 3.58-3.61 (m, 2H),4.43-4.45 (m, 2H), 4.74-4.78 (m, 1H), 6.78-6.81 (m, 1H), 6.90-6.92 (m,1H), 7.02-7.10 (m, 2H), 7.15-7.39 (m, 8H), 7.41 (d, 1H), 7.53 (s, 1H),7.55 (s, 1H) ppm.

MS (electrospray) m/z 755 [M+H]⁺

Preparation 57: 2-Hydroxy-1-naphthamide

A solution of 2-hydroxy-1-napthoic acid (5.0 g, 26.6 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.6 g, 29.2mmol), and 1-hydroxybenzotriazole (3.95 g, 29.2 mmol) in tetrahydrofuran(70 ml) was stirred at room temperature for 30 minutes prior to theaddition of 0.880 NH₃ (6 ml). The resulting suspension was stirred atroom temperature for 2 hours. The reaction mixture was filtered and thefiltrate diluted with water (80 ml) and extracted with ethyl acetate(4×80 ml). The combined organic extracts were washed with water (50ml×2), saturated aqueous sodium chloride (50 ml), dried (sodium sulfate)and the solvent removed in vacuo to give an orange oil. Purification bycolumn chromatography on silica gel eluting withdichloromethane:methanol:0.880 ammonia 95:5:0.5 gave the title compoundas a pink solid (1.83 g).

¹HNMR (400 MHz, CDCl₃): δ6.11-6.35 (bs, 2H), 7.17 (d, 1H), 7.36 (dd,1H), 7.54 (dd, 1H), 7.79 (d, 1H), 7.84 (d, 1H), 8.22 (d, 1H),11.70-11.88 (bs, 1H) ppm.

MS (electrospray) m/z 186 [M−H]⁻

Preparation 58: 6-Hydroxy-2-naphthamide

A solution of 6-hydroxy-2-napthoic acid (1.88 g, 9.99 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.11 g,10.98 mmol), 1-hydroxybenzotriazole (1.48 g, 10.98 mmol) and ammoniumcarbonate (4.80 g, 49.95 mmol) in N,N-dimethylformamide (70 ml) was leftto stir at room temperature under a nitrogen atmosphere for 3 days. Thesolvent was removed in vacuo and the residue partitioned betweensaturated aqueous sodium hydrogen carbonate (50 ml) and ethyl acetate(6×50 ml). The combined organic extracts were washed with water (25 ml),saturated aqueous sodium chloride (25 ml), dried (sodium sulfate) andthe solvent removed in vacuo. The solid was absorbed onto silica gel andpurified by column chromatography on silica gel eluting withdichloromethane:methanol:0.880 ammonia (95:5:0.5 changing to 90:10:1) togive the title compound as a pale yellow solid (1.1 g).

¹HNMR (400 MHz, CD₃OD) δ: 7.14 (d, 1H), 7.15 (s, 1H), 7.79 (d, 1H), 7.83(d, 2H), 8.32 (s, 1H) ppm.

MS (electrospray) m/z 186 [M−H]⁻

Preparation 59: 1-(Aminomethyl)-2-naphthol

A solution of borane in tetrahydrofuran (19.23 ml of a 1M solution,19.23 mmol) was added dropwise to a solution of the amide frompreparation 57 (0.90 g, 4.81 mmol) in tetrahydrofuran (10 ml) under anitrogen atmosphere. The reaction was then heated to reflux for 2 hours.The solution was cooled, treated with 6M hydrochloric acid (10 ml) andrefluxed for a further 2 hours. The resulting suspension was cooled andthe pH adjusted to pH9 by addition of 0.880 NH₃ and extracted with ethylacetate (50 ml×3). The combined organic extracts were washed with sat.aq. sodium chloride (20 ml), dried (sodium sulfate) and the solventremoved under reduced pressure. Purification by column chromatography onsilica gel eluting with dichloromethane:methanol:0.880:ammonia (95:5:0.5changing to 90:10:1) gave the title compound as a pink solid (0.19 g).

¹HNMR (400 MHz, CD₃OD) δ: 4.41 (s, 2H), 7.07 (d, 1H), 7.23 (1H, dd),7.43 (dd, 1H), 7.66 (d, 1H), 7.72 (d, 1H), 7.87 (d, 1H) ppm.

MS (electrospray) m/z 174 [M+H]⁺, 172 [M−H]⁻

Preparation 60: 6-(Aminomethyl)-2-naphthol

Prepared according to the method for preparation 59 using the amide frompreparation 58.

¹HNMR (400 MHz, CD₃OD) δ: 3.91 (s, 2H), 7.03-7.08 (m, 2H), 7.36 (dd,1H), 7.61 (d, 1H), 7.66 (d, 1H), 7.69 (s, 1H) ppm.

MS (electrospray) m/z 172 [M−H]⁻

Preparation 61: tert-Butyl (3-iodobenzyl)carbamate

A suspension of 3-iodobenzylamine hydrochloride (4.95 g, 18.4 mmol) indichloromethane (100 ml) was treated with triethylamine (3.1 ml, 22mmol) and di-t-butyl dicarbonate (4.40 g, 20 mmol) and the resultingsolution left to stir at room temperature under a nitrogen atmospherefor 1.5 hours. The reaction mixture was washed with 2M hydrochloric acid(30 ml), water (30 ml), dried (sodium sulfate), and the solvent removedin vacuo to give the title compound as a colourless solid (6.43 g).

¹HNMR (400 MHz, CDCl₃) δ: 1.46 (s, 9H), 4.21-4.30 (m, 2H). 4.79-4.89(bs, 1H), 7.06 (dd, 1H), 7.25 (d, 1H), 7.60 (d, 1H), 7.63 (s, 1H) ppm.

MS (electrospray) m/z 332 [M−H]⁻, 356 [M+Na]⁺

Preparation 62: tert-Butyl (2-bromobenzyl)carbamate

Prepared from 2-bromobenzylamine using the method of preparation 61 togive the title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD) δ: 1.50 (s, 9H), 4.33 (s, 2H), 7.18-7.22 (m, 1H),7.35-7.38 (m, 2H). 7.59 (d, 1H) ppm.

MS (electrospray) m/z 308/310 [M+Na]⁺

Preparation 63: tert-Butyl [(4′-hydroxybiphenyl-3-yl)methyl]carbamate

A solution of the iodide from preparation 61 (0.75 g, 2.25 mmol),4-hydroxy phenylboronic acid (0.629, 4.50 mmol),1,1′-bis(diphenylphosphino)ferrocenyl palladium(II) chloride (0.11 g,0.14 mmol), in N,N-dimethylformamide (14 ml) was treated with 2M aqueoussodium carbonate (4 ml) and the resulting mixture heated at 80° C. undera nitrogen atmosphere for 16 hours. The solvent was removed in vacuo andthe residue purified by column chromatography on silica gel eluting withethyl acetate:pentane (1:3) to give the title compound as a pale pinkcrystalline solid (0.73 g).

¹HNMR (400 MHz, CDCl₃) δ: 1.47 (s, 9H), 4.33-4.41 (m, 2H), 4.87-4.94(bs, 1H), 6.89 (d, 2H), 7.21 (d, 1H), 7.37 (dd, 1H), 7.43-7.45 (m, 4H)ppm.

MS (electrospray) m/z 298 [M−H]⁻, 322 [M+Na]⁺

Preparation 64: tert-Butyl [(2′-hydroxybiphenyl-3-yl)methyl]carbamate

Prepared from the iodide of preparation 61 and 2-hydroxyboronic acidusing the method of preparation 63 to give the title compound as acolourless solid.

¹HNMR (400 MHz, CDCl₃) δ: 1.46 (s, 9H), 4.38 (d, 2H), 4.90 (bs, 1H),5.24 (bs, 1H), 6.97-7.01 (m, 2H), 7.22-7.47 (m, 6H) ppm.

MS (electrospray) m/z 322 [M+Na]⁺

Preparation 65: tert-Butyl [(2′-hydroxybiphenyl-2-yl)methyl]carbamate

Prepared from the bromide of preparation 62 and 2-hydroxyboronic acidusing the method of preparation 63 to give the title compound as acolourless solid.

¹HNMR (400 MHz, CD₃OD) δ: 1.46 (s, 9H), 4.15 (d, 2H), 6.91-6.96 (m, 2H),7.10 (dd, 1H), 7.17-7.41 (m, 5H) ppm.

MS (electrospray) m/z 298 [M−H]⁻

Preparation 66: tert-Butyl [(3′-hydroxybiphenyl-2-yl)methyl]carbamate

Prepared from the bromide of preparation 62 and 3-hydroxyboronic acidusing the method of preparation 63 to give the title compound as acolourless solid.

¹HNMR (400 MHz, CD₃OD) δ: 1.48 (s, 9H), 4.21 (s, 2H), 6.76-6.83 (m, 3H),7.21-7.43 (m, 5H) ppm.

MS (electrospray) m/z 298 [M−H]⁻

Preparation 67: tert-Butyl [(3′-hydroxybiphenyl-3-yl)methyl]carbamate

Prepared from the iodide of preparation 61 and 3-hydroxy phenylboronicacid using the method of preparation 63 to give the title compound as abrown gum.

¹HNMR (400 MHz, CDCl₃) δ: 1.48 (s, 9H), 4.37 (d, 2H), 4.86-4.91 (bs,1H), 6.82 (dd, 1H), 7.04 (t, 1H), 7.11 (d, 1H), 7.24-7.30 (m, 2H), 7.36(t, 1H), 7.43 (d, 1H), 7.45 (d, 1H) ppm.

MS (electrospray) m/z 298 [M−H]⁻, 597 [2M−H]⁻

Preparation 68: 3′-(Aminomethyl)biphenyl-4-ol

The phenol from preparation 63 (0.73 g, 2.43 mmol) was treated with 4MHCl in dioxan (6 ml, 24.3 mmol) and the resulting solution allowed tostir at room temperature for 3 hours. The solvent was removed in vacuoto give the title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD) δ: 4.17 (s, 2H), 6.87 (d, 2H), 7.34 (d, 1H),7.45-7.50 (m, 3H), 7.61 (d, 1H), 7.65 (s, 1H) ppm.

MS (electrospray) m/z 198 [M−H]⁻, 200 [M+H]⁺

Preparation 69: 3′-(Aminomethyl)biphenyl-3-ol

Prepared from the phenol of preparation 67 using the method ofpreparation 68 to give the title compound as a brown gum.

¹HNMR (400 MHz, CD₃OD) δ: 4.17 (s, 2H), 6.80 (dd, 1H), 7.04 (t, 1H),7.08-7.11 (m, 1H), 7.26 (t, 1H), 7.41 (d, 1H), 7.50 (t, 1H), 7.63 (d,1H), 7.69 (s, 1H) ppm.

MS (electrospray) m/z 198 [M−H]⁻, 200 [M+H]⁺

Preparation 70: 3′-(Aminomethyl)biphenyl-2-ol

Prepared from the phenol of preparation 63 using the method ofpreparation 68 to give the title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD) δ: 4.19 (s, 2H), 6.93-6.97 (m, 2H), 7.19-7.23 (m,1H), 7.31 (d, 1H), 7.41 (dd, 1H), 7.50-7.53 (m, 1H), 7.65-7.69 (m, 2H)ppm.

MS (electrospray) m/z 200 [M+H]⁺

Preparation 71: 2′-(Aminomethyl)biphenyl-2-ol

Prepared from preparation 65 using the method of preparation 68 to givethe title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD) δ:4.03 (s, 2H), 6.99-7.04 (m, 2H), 7.19 (dd, 1H),7.30-7.34 (m, 2H), 7.50-7.58 (m, 3H) ppm.

MS (electrospray) m/z 200 [M+H]⁺

Preparation 72: 2′-(Aminomethyl)biphenyl-3-ol

Prepared from preparation 66 using the method of preparation 68 to givethe title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD): 4.15 (s, 2H), 6.79-6.84 (m, 2H), 6.88-6.91 (m,1H), 7.31-7.35 (m, 1H), 7.37-7.40 (m, 1H), 7.48-7.54 (m, 2H), 7.56-7.60(m, 1H) ppm.

MS (electrospray) m/z 200 [M+H]⁺

Preparation 73: (4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)acetonitrile

A solution of (4-hydroxyphenyl)acetonitrile (6.01 g, 45.1 mmol) inN,N-dimethylformamide (60 ml) was treated with imidazole (3.819, 58.6mmol), tert-butyldimethylsilyl chloride (7.49 g, 49.6 mmol) andN,N-dimethylaminopyridine (20 mg) and the resulting solution left tostir at room temperature under a nitrogen atmosphere for 16 hours. Thereaction mixture was diluted with water (200 ml) and extracted withethyl acetate (200 ml×2). The combined organic extracts were washed withsat. aq. sodium chloride (200 ml), dried (sodium sulfate) and thesolvent removed in vacuo. Purification by column chromatography onsilica gel eluting with ethyl acetate:pentane (0:100 changing to 10:90)gave the title compound as a pale yellow oil (9.44 g).

¹HNMR (400 MHz, CDCl₃) δ: 0.19 (s, 6H), 0.97 (s, 9H), 3.66 (s, 2H), 6.82(d, 2H), 7.17 (d, 2H) ppm.

MS (APCI) m/z 265 [M+NH₄]⁺

Preparation 74: 2-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-2-methylpropanenitrile

A solution of the nitrile from preparation 73 (5.62 g, 22.7 mmol),methyl iodide (3.11 ml, 50 mmol), and 18-crown-6 (1.5 g, 5.6 mmol) indry tetrahydrofuran (300 ml) was cooled to −78° C. under a nitrogenatmosphere. Potassium tert-butoxide (50 ml of a 1 M solution intetrahydrofuran, 50 mmol) was added dropwise over 20 minutes and thereaction mixture then allowed to warm gradually to room temperature.After 2 hours the reaction was recooled to −78° C. and quenched byaddition of sat. aq. ammonium chloride (200 ml) and allowed to warm toroom temperature. The resulting solution was extracted with ethylacteate (300 ml×2), the combined organics were dried (sodium sulfate)and the solvent removed in vacuo. Purification by column chromatographyon silica gel eluting with ethyl acetate:pentane (0:100 changing to10:90) gave the title compound as a colourless oil (4.75 g).

¹HNMR (400 MHz, CDCl₃) δ: 0.19 (s, 6H), 0.97 (s, 9H), 1.68 (s, 6H), 6.82(d, 2H), 7.30 (d, 2H) ppm.

MS (APCI) m/z 293 [M+NH₄]⁺

Preparation 75: [2-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-2-methylpropyl]amine

A solution of the nitrile from preparation 74 (0.75 g, 2.7 mmol) indiethyl ether (5 ml) was added dropwise to a cold (0° C.) solution oflithium aluminium hydride in diethyl ether (2.98 ml of a 1 M solution).The resulting solution was stirred at 0° C. for 3 hours and thenquenched by addition of water (0.1 ml), 2N aqueous sodium chloride (0.1ml), and further water (0.3 ml). The resulting suspension was filteredand the filtrate concentrated in vacuo. Purification by columnchromatography on silica gel eluting with dichloromethane:methanol:0.880ammonia (97:3:0.5 changing to 93:7:0.5) gave the title compound as acolourless oil (0.52 g).

¹HNMR (400 MHz, CDCl₃) δ: 0.18 (s, 6H), 0.97 (s, 9H), 1.00 (bs, 2H),1.25 (s, 6H), 2.73 (s, 2H), 6.78 (d, 2H), 7.16 (d, 2H) ppm.

MS (APCI) m/z 280 [M+H]⁺

Preparation 76: 4-(Aminomethyl)-2,6-dimethylphenol hydrochloride

A solution of borane in tetrahydrofuran (27.1 ml of a 1M solution, 27.1mmol) was added dropwise to a solution of3,5-dimethyl-4-hydroxybenzonitrile (1.0 g, 6.79 mmol) in tetrahydrofuran(70 ml) and the resulting solution heated to reflux under a nitrogenatmosphere for 16 hours. The reaction was cooled to room temperature andtreated with 6N hydrochloric acid (20 ml) and refluxed for a further 30minutes. The reaction mixture was cooled to room temperature and thesolvent removed in vacuo. Purification using strong cation exchangeresin, eluting biproducts with methanol followed by 2M ammonia inmethanol to elute the product gave the title compound as an orange oil.The oil was treated with 1M hydrogen chrloide in methanol (20 ml) andthe solvent removed in vacuo to give the title compound as a pale yellowsolid (1.12 g).

¹HNMR (400 MHz, CDCl₃) δ: 2.22 (s, 6H), 3.75 (s, 2H), 6.90 (s, 2H).

Preparation 77: 2-(Aminomethyl)-4-chlorophenol hydrochloride

Prepared from 5-chloro-2-hydroxybenzonitrile using the proceduredescribed in preparation 76.

¹HNMR (400 MHz, CDCl₃) δ: 4.08 (s, 2H), 6.87 (d, 1H), 7.27 (d, 1H), 7.35(s, 1H).

MS (APCI) m/z 156 [M−H]⁻, 158 [M+H]⁺

Preparation 77: 4′-(Aminomethyl)biphenyl-4-ol hydrochloride

Prepared from 4′-hydroxybiphenyl-4-carbonitrile using the proceduredescribed in preparation 76.

¹HNMR (400 MHz, CD₃OD) δ: 4.10 (s, 2H), 6.83 (d, 2H), 7.44-7.46 (m, 4H),7.60 (d, 2H) ppm.

Preparation 79: 3,5-Dichloro-N-ethyl-2-hydroxybenzamide

Prepared from 3,5-dichloro-2-hydroxybenzoic acid and ethylamine usingthe method of preparation 57 to give the title compound as a pale yellowsolid.

¹HNMR (400 MHz, CDCl₃) δ: 1.28 (t, 3H), 3.47-3.54 (m, 2H), 6.29-6.36(bs, 1H), 7.27 (d, 1H), 7.48 (d, 1H) ppm.

MS (electrospray) m/z 232 [M−H]³¹

Preparation 80: 2,4-Dichloro-6-[(ethylamino)methyl]phenol

A solution of the amide from preparation 79 (0.77 g, 3.29 mmol) intetrahydrofuran (10 ml) was cooled to 0° C. and treated withborane.tetrahydrofuran complex (9.9 ml of a 1M solution intetrahydrofuran, 9.9 mmol). The resulting solution was allowed to warmto room temperature over 20 minutes and then heated to relux for 16hours. The reaction mixture was cooled to 0° C. and quenched by additionof methanol (until effervescence ceased). The resulting solution wasallowed to warm to room temperature over 2 hours and then the solventwas removed in vacuo. The residue was dissolved in dichloromethane (40ml) and washed with water (10 ml×2), sat. aq. sodium chloride (10 ml),dried (sodium sulfate) and reduced in vacuo to give a colourless oil.Purification by column chromatography on silica gel eluting withdichloromethane:methanol (98:2 changing to 95:5) gave the title compoundas a colourless solid (0.53 g).

¹HNMR (400 MHz, CDCl₃) δ: 1.17 (t, 3H), 2.72 (q, 2H), 3.98 (s, 2H), 6.86(d, 1H), 7.23 (d, 1H) ppm.

Preparation 81: 6-Hydroxy-N-methyl-1-naphthamide

Prepared from 6-hydroxy-1-naphthoic acid and methylamine using themethod of preparation 57 to give the title compound as a pale orangesolid.

¹HNMR (400 MHz, CD₃OD) δ: 2.97 (s, 3H), 7.10-7.14 (m, 2H), 7.34-7.40 (m,2H), 7.73 (dd, 1H), 8.04 (d, 1H) ppm.

Preparation 82: 5-[(Methylamino)methyl]-2-naphthol

Prepared from the amide of preparation 81 using the method ofpreparation 80 to give the title compound as a pale pink solid.

¹HNMR (400 MHz, CD₃OD) δ: 2.48 (s, 3H), 4.14 (s, 2H), 7.11-7.14 (m, 2H),7.25 (d, 1H), 7.33 (t, 1H), 7.59 (d, 1H), 7.94 (d, 1H) ppm.

MS (electrospray) m/z 186 [M−H]⁻, 188 [M+H]⁺

Preparation 83: 3-Hydroxy-5-(trifluoromethyl)benzamide

Prepared from 3-hydroxy-5-(trifluoromethyl)benzoic acid using the methodof preparation 58 to give the title compound as a pale yellow solid.

¹HNMR (400 MHz, CD₃OD) δ: 7.18 (t, 1H), 7.50 (t, 1H), 7.60-7.61 (m, 1H)ppm.

MS (electrospray) m/z 204 [M−H]⁻

Preparation 84: 3-(Aminomethyl)-5-(trifluoromethyl)phenol

Prepared from the amide of preparation 83 using the method ofpreparation 80 to give the title compound as a pale yellow oil.

¹HNMR (400 MHz, CD₃OD) δ: 3.81 (s, 2H), 6.91 (s, 1H), 6.98 (s, 1H), 7.09(s, 1H) ppm.

MS (electrospray) m/z 192 [M+H]⁺

Preparation 85: 3-(Aminomethyl)-5-chlorophenol

Prepared from 3-chloro-5-hydroxybenzonitrile using the method ofpreparation 76 to give the title compound as a pale yellow solid.

¹HNMR (400 MHz, CD₃OD) δ:3.69 (s, 2H), 6.65 (d, 2H), 6.79 (t, 1H) ppm.

MS (electrospray) m/z 158 [M+H]⁺

Preparation 86: 3-[(Acetylamino)methyl]-5-chlorophenyl acetate

A solution of the amine of preparation 85 (700 mg, 4.46 mmol) intetrahydrofuran (20 ml) was treated with triethylamine (1.3 ml, 8.9mmol) and acetyl chloride (0.64 ml, 8.9 mmol). The resulting mixture wasleft to stir at room temperature for 1 hour. The reaction mixture wasfiltered and the filtrate reduced in vacuo to give the title compound asa colourless solid (1.07 g).

¹HNMR (400 MHz, CDCl₃) δ: 2.15 (s, 3H), 2.27 (s, 3H), 3.71-3.75 (m, 1H),4.38-4.41 (m, 2H), 6.92 (s, 1H), 7.02 (s, 1H), 7.13 (s, 1H) ppm.

MS (electrospray) m/z 264 [M+Na]⁺

Preparation 87: N-(3-Chloro-5-hydroxybenzyl)acetamide

A solution of the diacetate of preparation 86 (1.07 g, 4.44 mmol) inmethanol (10 ml) was treated with sodium methoxide (30 mg, 0.55 mmol)and the resulting mixture left to stir at room temperature for 6 hours.The solvent was removed in vacuo and the residue purified by columnchromatography on silica gel eluting with ethyl acetate:pentane (1:1changing to 1:0) gave the title compound as a yellow solid (0.78 g).

¹HNMR (400 MHz, CDCl₃) δ: 2.05 (s, 3H), 4.33 (d, 2H), 6.08-6.14 (m, 1H),6.73 (d, 2H), 6.79 (t, 1H) ppm.

MS (electrospray) m/z 200 [M+H]⁺

Preparation 88: 3-Chloro-5-[(ethylamino)methyl]phenol

Prepared from the amide of preparation 87 (0.75 g, 3.76 mmol)) using themethod of preparation 59 to give the title compound as a colourlesssolid (0.48 g).

¹HNMR (400 MHz, CD₃OD) δ: 1.14 (t, 3H), 2.71 (q, 2H), 3.25-3.27 (m, 1H),3.72 (s, 2H), 6.66-6.68 (m, 2H), 6.79 (s, 1H) ppm.

MS (electrospray) m/z [M−H]⁻

Preparation 89: 4-([tert-Butyl(dimethyl)silyl]oxy)-2-chlorobenzaldehyde

A solution of 2-chloro-4-hydroxybenzaldehyde (5.0 g, 32 mmol),tert-butyl(dimethyl)silyl chloride (5.3 g, 35 mmol), imidazole (2.9 g,45 mmol) and N,N-dimethylaminopyridine (10 mg) in N,N-dimethylformamide(40 ml) was stirred at room temperature under a nitrogen atmosphere for16 hours. The solvent was removed in vacuo and the residue partitionedbetween ethyl acteate (100 ml) and water (100 ml). The organic phase wasseparated, washed with sat. aq. sodium chloride (50 ml), dried (sodiumsulfate) and reduced in vacuo. Further purification by columnchromatography on silica gel eluting with pentane:ethyl acetate (3:1changing to 2:1) gave the title compound as a colourless oil (6.50 g).

¹HNMR (400 MHz, CDCl₃) δ: 0.25 (s, 6H), 0.97 (s, 9H), 6.80 (dd, 1H),6.87 (d, 1H), 7.84 (d, 1H), 10.32 (s, 1H) ppm.

Preparation 90:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}-2-chlorobenzyl)prop-2-en-1-amine

A solution of the aldehyde from preparation 89 (6.50 g, 24.0 mmol) andallylamine (1.51 g, 26.4 mmol) in dichloromethane (60 ml) was treatedwith sodium triacetoxyborohydride (7.6 g, 35.6 mmol) and the resultingsuspension allowed to stir at room temperature for 16 hours. Sat. aq.sodium bicarbonate (50 ml) was added and the organic phase separated.The organic phase was washed with sat. aq. sodium chloride (50 ml),dried (sodium sulfate) and the solvent removed in vacuo to give a yellowoil. Purification by column chromatography on silica gel eluting withpentane:ethyl acetate (3:1 changing to 2:1) gave the title compound as acolourless oil (2.80 g).

¹HNMR (400 MHz, CDCl₃) δ: 0.19 (s, 6H), 0.97 (s, 9H), 1.84 (bs, 1H),3.26 (d, 2H), 3.81 (s, 2H), 5.12 (dd, 1H), 5.20 (dd, 1H), 5.88-5.98 (m,1H), 6.71 (dd, 1H), 6.85-6.86 (d, 1H), 7.24 (d, 1H) ppm.

MS (electrospray) m/z 312 [M+H]⁺

Preparation 91: 4-[(Allylamino)methyl]-2,6-dichlorophenol

Prepared from 3,5-dichloro-4-hydroxybenzaldehyde and allylamine usingthe method of preparation 90 to give the title compound as a colourlessoil.

¹HNMR (400 MHz, DMSOd₆) δ: 3.11 (d, 2H), 3.50 (s, 2H), 5.06 (d, 1H),5.16 (d, 1H), 5.77-5.90 (m, 1H), 7.10 (s, 2H) ppm.

MS (electrospray) m/z 232/234 [M+H]⁺

Preparation 92: (4-{[tert-Butyl(dimethyl)silyl]oxy}-2-chlorobenzyl)amine

A solution of the amine from preparation 91 (2.8 g, 9.0 mmol), dimethylbarbituric acid (7.0 g, 45 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.10 g, 0.08 mmol) in dichloromethane (80 ml) was heatedto reflux for 4 hours. The cooled solution was reduced in vacuo and theresidue partitioned between ethyl acetate (50 ml) and 1N aqueous sodiumhydroxide (50 ml). The organic phase was separated, washed with sat. aq.sodium chloride (50 ml), dried (sodium sulfate) and reduced in vacuo.Further purification by column chromatography on silica gel eluting withdichloromethane:methanol:0.880 ammonia (98:2:0 changing to 95:5:0.5)gave the title compound as a colourless oil (1.70 g).

¹HNMR (400 MHz, CDCl₃) δ: 0.19 (s, 6H), 0.97 (s, 9H), 1.89 (s, 2H), 3.85(s, 2H), 6.70 (dd, 1H), 6.85-6.86 (dd, 1H), 7.21 (d, 1H) ppm.

Preparation 93: (4-{[tert-Butyl(dimethyl)silyl]oxy}benzyl)methylamine

Prepared from the aldehyde of preparation 89 and methylamine using themethod of preparation 90 to give the title compound as a yellow oil.

¹HNMR (400 MHz, CDCl₃) δ: 0.23 (s, 6H), 1.00 (s, 9H), 2.50 (s, 3H), 3.93(s, 2H), 6.70-6.73 (m, 1H), 6.76 (d, 1H), 7.20 (d, 1H) ppm.

MS (electrospray) m/z 286/288 [M+H]⁺

Preparation 94: 4-(Aminomethyl)-2,6-dichlorophenol barbiturate

Prepared from the amine from preparation 91 using the method ofpreparation 92 to give the title compound as the barbituric acid salt.

¹HNMR (400 MHz, DMSOd₆) δ: 2.60-4.40 (broad multiplet, 2H), 3.03 (s,6H), 3.93 (s, 2H), 7.49 (s, 2H) ppm.

MS (electrospray) m/z 192/194[M+H]⁺

Preparation 95: 4-{[tert-Butyl(dimethyl)silyl]oxy}-2,3-dichlorobenzaldehyde

Prepared from 2,3-dichloro-4-hydroxybenzaldehyde according to the methodfor preparation 89 to give the title compound as yellow oil.

¹HNMR (400 MHz, CDCl₃) δ: 0.29 (s, 6H), 1.04 (s, 9H), 6.88 (d, 1H), 7.76(d, 1H), 10.32 (s, 1H) ppm.

Preparation 96:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}-2,3-dichlorobenzyl)prop-2-en-1-amine

Prepared according to preparation 90 using allylamine and the aldehydefrom preparation 95 to give the title compound as a colourless oil.

¹HNMR (400 MHz, CDCl₃) δ: 0.20 (s, 6H), 1.01 (s, 9H), 3.25 (d, 2H), 3.82(s, 2H), 5.10 (dd, 1H), 5.18 (dd, 1H), 5.85-5.93 (m, 1H), 6.76 (d, 1H),7.13 (d, 1H) ppm.

MS (electrospray) m/z 346/348 [M+H]⁺

Preparation 97:(4-{[tert-Butyl(dimethyl)silyl]oxy}-2,3-dichlorobenzyl)amine

Prepared according to preparation 92 using amine from preparation 96 togive the title compound as a colourless oil.

¹HNMR (400 MHz, CD₃OD) δ: 0.23 (s, 6H), 1.03 (s, 9H), 3.92 (s, 2H), 6.77(d, 1H), 7.12 (d, 1H) ppm.

Preparation 98: 4-{[tert-Butyl(dimethyl)silyl]oxy}-1-naphthaldehyde

Prepared from 2,3-dichloro-4-hydroxybenzaldehyde according to the methodfor preparation 89 to give the title compound as brown solid.

¹HNMR (400 MHz, CDCl₃) δ: 0.36 (s, 6H), 1.10 (s, 9H), 6.94 (d, 1H), 7.56(dd, 1H), 7.68 (dd, 1H), 7.86 (d, 1H), 8.27 (dd, 1H), 9.30 (dd, 1H),10.21 (s, 1H) ppm.

Preparation 99:N-[(4-{[tert-Butyl(dimethyl)silyl]oxy}-1-naphthyl)methyl]prop-2-en-1-amine

Prepared according to preparation 90 using allylamine and the aldehydefrom preparation 98 to give the title compound as a yellow oil.

¹HNMR (400 MHz, CDCl₃) δ: 0.30 (s, 6H), 1.11 (s, 9H), 1.97 (bs, 1H),3.39 (d, 2H), 4.17 (s, 2H), 5.16 (dd, 1H), 5.25 (dd, 1H), 5.95-6.05 (m,1H), 6.82 (d, 1H), 7.32 (d, 1H), 7.47-7.57 (m, 2H), 8.07 (d, 1H), 8.25(d, 1H) ppm.

MS (electrospray) m/z 328 [M+H]⁺, 655 [2M+H]⁺

Preparation 100:[(4-{[tert-butyl(dimethyl)silyl]oxy}-1-naphthyl)methyl]amine

Prepared according to preparation 92 using amine from preparation 99 togive the title compound as a colourless oil.

¹HNMR (400 MHz, CDCl₃) δ: 0.28 (s, 6H), 1.09 (s, 9H), 2.31 (bs, 2H),4.24 (s, 2H), 6.80 (d, 1H), 7.27 (t, 1H), 7.46-7.55 (m, 4H), 8.00 (d,1H), 8.25 (d, 1H).

Preparation 101: 3-Hydroxy-N-methyl-5-(trifluoromethyl)benzamide

Prepared from 3-hydroxy-5-(trifluoromethyl)benzoic acid and methylamineusing the method of preparation 58 to give the title compound as a paleorange solid.

¹HNMR (400 MHz, CD₃OD) δ: 2.99 (s, 3H), 7.14 (s, 1H), 7.43 (s, 1H), 7.52(s, 1H) ppm.

MS (electrospray) m/z 218 [M−H]⁻

Preparation 102: 3-[(Methylamino)methyl]-5-(trifluoromethyl)phenol

Prepared from the amide of preparation 101 using the method ofpreparation 59 to give the title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD) δ: 2.41 (s, 3H), 3.75 (s, 2H), 6.93 (s, 1H), 6.98(s, 1H), 7.09 (s, 1H) ppm.

MS (electrospray) m/z 206 [M+H]⁺

Preparation 103: 4-(Aminomethyl)-3,5-dimethylphenol

Prepared from 4-hydroxy-2,6-dimethylbenzonitrile using the method ofpreparation 76 to give the title compound as a colourless solid.

¹HNMR (400 MHz, D₂O) δ: 2.09 (s, 6H), 3.90 (s, 2H), 6.95 (s, 2H) ppm.

Preparation 104: (4-Hydroxy-2,5-dimethylphenyl)acetonitrile

A solution of (4-methoxy-2,5-dimethylphenyl)acetonitrile (0.5 g, 2.9mmol) in dichloromethane (10 ml) was cooled to −80° C. and treated witha solution of boron tribromide in dichloromethane (14.3 ml of a 1Msolution, 14.3 mmol). The reaction mixture was stirred at −80° C. for afurther 30 minutes and then gradually allowed to warm to roomtemperature over a period of 2 hours. The reaction mixture was quenchedwith saturated aqueous sodium bicarbonate (20 ml) and the organic phaseseparated. The organic phase was washed with saturated aqueous sodiumchloride (20 ml), dried (sodium sulfate) and the solvent removed invacuo to give a pale brown solid. Purification by column chromatographyon silica gel eluting with ethyl acetate:pentane (1:4 changing to 1:2)gave the title compound as a colourless solid (0.28 g).

¹HNMR (400 MHz, CD₃OD) δ: 2.13 (s, 3H), 2.23 (s, 3H), 3.66 (s, 2H), 6.60(s, 1H), 6.98 (s, 1H) ppm.

MS (electrospray) m/z 160 [M−H]⁻

Preparation 105: (4-Hydroxy-2,3-dimethylphenyl)acetonitrile

Prepared from (4-methoxy-2,3-dimethylphenyl)acetonitrile using themethod from preparation 104 to give the title compound as a pale yellowsolid.

¹HNMR (400 MHz, CDCl₃) δ: 2.20 (s, 3H), 2.24 (s, 3H), 3.62 (s, 2H), 4.91(bs, 1H), 6.64 (d, 1H), 7.03 (d, 1H) ppm.

MS (electrospray) m/z 160 [M−H]⁻

Preparation 106: (4-Hydroxy-3-methylphenyl)acetonitrile

Prepared from (4-methoxy-3-methylphenyl)acetonitrile using the methodfrom preparation 104 to give the title compound as a pale yellow solid.

¹HNMR (400 MHz, CDCl₃) δ: 2.25 (s, 3H), 3.65 (s, 2H), 4.98 (bs, 1H),6.76 (d, 1H), 7.01 (d, 1H), 7.07 (s, 1H) ppm.

MS (electrospray) m/z 146 [M−H]⁻

Preparation 107: 4-(2-Aminoethyl)-2,5-dimethylphenol

A solution of the nitrile from preparation 104 (0.28 g, 1.74 mmol) inethanol (15 ml) was hydrogenated at 60 psi over Raney Nickel (0.1 g, 50%w/w) for 16 hours. The reaction mixture was filtered and the solventremoved in vacuo. The residue was purified by strong cation exchangeresin eluting non-basic impurities with methanol and then 1M ammonia inmethanol to give the title compound as a colourless oil.

¹HNMR (400 MHz, CD₃OD) δ: 2.11 (s, 3H), 2.19 (s, 3H), 2.63-2.67 (m, 2H),2.72-2.76 (m, 2H), 6.54 (s, 1H), 6.81 (s, 1H) ppm.

MS (electrospray) m/z 166 [M+H]⁺

Preparation 108: 4-(2-Aminoethyl)-2,3-dimethylphenol

Prepared from the nitrile of preparation 105 using the method ofpreparation 107 to give the title compound as a colourless oil.

¹HNMR (400 MHz, CD₃OD) δ: 2.12 (s, 3H), 2.19 (s, 3H), 2.68-2.75 (m, 4H),6.55 (d, 1H), 6.78 (d, 1H) ppm.

MS (electrospray) m/z 166 [M+H]⁺

Preparation 109: 4-(2-Aminoethyl)-2-methylphenol

Prepared from the nitrile of preparation 106 using the method ofpreparation 107 to give the title compound as a colourless oil.

¹HNMR (400 MHz, CD₃OD) δ: 2.15 (s, 3H), 2.60-2.64 (m, 2H), 2.79-2.83 (m,2H), 6.66 (d, 1H), 6.82 (d, 1H), 6.90 (s, 1H) ppm.

MS (electrospray) m/z 152 [M+H]⁺

EXAMPLES 1-38

The appropriate protected alcohol (0.075 mmol) was dissolved in ethanol(4 ml) and the solution treated with a solution of ammonium fluoride (16mg, 0.43 mmol) in water (300 μL). The reaction mixture was then stirredat 50° C. for 18 hours before being allowed to cool to room temperature.If a solid product precipitated the reaction mixture was filtered andwashed with methanol:water (2 ml, 1:1 by volume) to give the titlecompound. If no product precipitated the reaction mixture wasconcentrated in vacuo and the residue purified by column chromatographyon silica gel eluting with dichloromethane:methanol:0.88 ammonia 98:2:0to 95:5:0.5 to 90:10:1 to yield the title product.

Alternatively, the following process can be used for the synthesis ofexamples 1 to 38: A solution of the appropriate protected alcohol (2.87mmol) in methanol (80 ml) is treated with a solution of ammoniumfluoride (1.06 g, 28.7 mmol) in water (53 ml) and the resulting mixtureheated at 40° C. for 16 hours. The reaction is cooled to roomtemperature and filtered, washing with a mixture of water and methanol(1:1 by volume, 3×10 ml), methanol (2×10 ml). The solid is dried invacuo to give the desired compound.

EXAMPLE 12-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-(4-hydroxy-3-methoxybenzyl)acetamide

Preparation 18 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was concentrated in vacuo and the residue purified by columnchromatography on silica gel eluting with dichloromethane:methanol:0.88ammonia 98:2:0 to 95:5:0.5 to 90:10:1 to yield the title product as acolourless solid.

¹HNMR (CD₃OD, 400 MHz) δ: 1.04 (s, 3H), 1.06 (s, 3H), 2.68-2.90 (m, 7H),3.53 (s, 2H), 3.74 (s, 3H), 4.23 (m, 2H), 4.62 (m, 1H), 6.67 (m, 2H),6.77 (m, 1H), 6.85 (d, 1H), 7.01-7.22 (m, 6H), 7.37 (m, 1H) ppm.

MS (electrospray) m/z 572 [M+H]⁺

EXAMPLE 2N-[(4′-Hydroxybiphenyl-4-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide

Preparation 19 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was filtered and the solid washed with methanol:water (2 ml, 1:1by volume) to give the title compound as a colourless solid.

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.90 (s, 3H), 0.92 (s, 3H), 2.56 (s, 2H),2.62-2.65 (m, 2H), 2.88 (s, 3H), 3.43 (s, 2H), 4.25 (2H, d), 4.40-4.43(m, 1H), 6.80-6.82 (m, 3H), 6.96-7.01 (m, 2H), 7.07-7.10 (m, 2H),7.14-7.18 (m, 2H), 7.23 (d, 2H), 7.42-7.48 (m, 4H), 8.47 (t, 1H).

MS (electrospray) m/z 618 [M+H]⁺

EXAMPLE 3N-(4-Chloro-2-hydroxybenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide

Preparation 20 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was filtered and washed with methanol:water (2 ml, 1:1 byvolume) to give the title compound as a colourless solid.

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.90 (s, 3H), 0.91 (s, 3H), 2.56 (s, 2H),2.59-2.67 (m, 2H), 2.88 (s, 3H), 3.44 (s, 2H), 4.16 (s, 2H), 4.40-4.43(m, 1H), 6.76-6.81 (m, 2H), 6.96-7.18 (m, 8H), 8.42 (s, 1H).

MS (electrospray) m/z 574 [M−H]⁻, 576 [M+H]⁺

EXAMPLE 4N-(4-Hydroxy-3,5-dimethylbenzyl)-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide

Preparation 21 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was filtered and washed with methanol:water (2 ml, 1:1 byvolume) to give the title compound as a colourless solid.

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.90 (s, 3H), 0.91 (s, 3H), 2.08 (s, 6H),2.55 (s, 2H), 2.62-2.65 (m, 2H), 2.88 (s, 3H), 3.38 (s, 2H partiallyobscured by H₂O), 4.05 (d, 2H), 4.40-4.43 (m, 1H), 6.71 (s, 2H), 6.81(d, 1H), 6.95-7.01 (m, 2H), 7.05-7.09 (m, 2H), 7.13-7.18 (m, 2H),8.28-8.31 (t, 1H).

EXAMPLE 52-(3-{2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)-N-[(2-hydroxy-1-naphthyl)methyl]acetamide

Preparation 22 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 L). The reaction mixture was then stirred at 50° C. for 18hours before being allowed to cool to room temperature. The reactionmixture was concentrated in vacuo and the residue purified by columnchromatography on silica gel eluting with dichloromethane:methanol:0.88ammonia 98:2:0 to 95:5:0.5 to 90:10:1 to yield the title product as acolourless solid.

¹HNMR (400 MHz, DMSOd₆) δ: 0.86 (s, 3H), 0.87 (s, 3H), 2.46-2.68 (m,4H), 2.90 (s, 3H), 3.40 (s, 2H), 4.41-4.47 (m, 1H), 4.63 (d, 2H), 6.83(d, 1H), 6.94-7.05 (m, 4H), 7.11-7.16 (m, 2H), 7.19 (s, 1H), 7.27 (t,1H), 7.40 (t, 1H), 7.72 (d, 1H), 7.79 (d, 1H), 7.88 (d, 1H), 8.48-8.52(bs, 1H).

MS (electrospray) m/z 590 [M−H]⁻

EXAMPLE 62-(3-(2-[((2R)-2-Hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl)phenyl)-N-[(6-hydroxy-2-naphthyl)methyl]acetamide

Preparation 23 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was filtered and washed with methanol:water (2 ml, 1:1 byvolume) to give the title compound as a colourless solid.

¹HNMR (400 MHz, DMSO-d₆) δ: 0.90 (s, 3H), 0.92 (s, 3H), 2.49-2.68 (m,4H), 2.89 (s, 3H), 3.44 (s, 2H), 4.34 (d, 2H), 4.40-4.43 (m, 1H), 6.80(d, 1H), 6.96-7.17 (m, 7H), 7.23 (d, 1H), 7.51 (s, 1H), 7.58 (d. 1H),7.61 (d, 1H), 8.50 (dd, 1H).

MS (electrospray) m/z 590 [M−H]⁻, 592 [M+H]⁺, 614 [M+Na]⁺

EXAMPLE 7N-[(4′-Hydroxybiphenyl-3-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide

Preparation 24 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was filtered and washed with methanol:water (2 ml, 1:1 byvolume) to give the title compound as a colourless solid.

¹HNMR (400 MHz, DMSOde) δ: 0.88 (s, 3H), 0.90 (s, 3H), 2.66-2.54 (m,4H), 2.88 (s, 3H), 3.43 (s, 2H), 4.29 (d, 2H), 4.39-4.43 (m, 1H),6.79-6.82 (m, 3H), 6.96-7.01 (dd, 2H), 7.07-7.17 (m, 5H), 7.27-7.31 (dd,1H), 7.36-7.41 (m, 4H), 8.52 (dd, 1H) ppm.

MS (electrospray) m/z 618 [M+H]⁺

EXAMPLE 8N-[(3′-Hydroxybiphenyl-3-yl)methyl]-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide

Preparation 25 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was filtered and washed with methanol:water (2 ml, 1:1 byvolume) to give the title compound as a colourless solid.

¹HNMR (400 MHz, DMSOde) δ: 1.16 (s, 6H), 2.85 (s, 2H), 2.92 (s, 3H),2.96-3.03 (m, 2H), 3.57 (s, 2H), 4.42 (s, 2H), 4.77-4.79 (m, 1H), 6.74(d, 1H), 6.90 (d, 1H), 6.95-6.97 (m, 2H), 7.09-7.27 (m, 7H), 7.32 (t,1H), 7.41-7.42 (m, 3H) ppm.

MS (electrospray) m/z 618 [M+H]⁺, 6401[M+Na]⁺, 616 [M−H]⁻

EXAMPLE 92-(3-{2-[((2R)-2-Hydroxy-2-(4-hydroxy-3-[(methylsulfonyl)amino]phenyl)ethyl)amino]-2-methylpropyl}phenyl)-N-[2-(4-hydroxyphenyl)-2-methylpropyl]acetamide

Preparation 26 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was concentrated in vacuo and the residue purified by columnchromatography on silica gel eluting with dichloromethane:methanol:0.88ammonia 98:2:0 to 95:5:0.5 to 90:10:1 to yield the title product as acolourless solid.

¹HNMR (400 MHz, DMSOd₆) δ: 0.91 (s, 3H), 0.92 (s, 3H), 1.11 (s, 6H),2.56 (s, 2H), 2.64-2.66 (m, 2H), 2.89 (s, 3H), 3.15 (s, 2H), 3.35 (s,2H), 4.42-4.45 (m, 1H), 6.65 (d, 2H), 6.81 (d, 1H), 6.94-7.03 (m, 4H),7.07-7.14 (m, 3H), 7.18 (s, 1H), 7.60 (t, 1H).

MS (APCI) m/z 582 [M−H]⁻, 584 [M+H]⁺

EXAMPLE 10N-(3,5-Dichloro-2-hydroxybenzyl)-N-ethyl-2-(3-{2-[((2R)-2-hydroxy-2-{4-hydroxy-3-[(methylsulfonyl)amino]phenyl}ethyl)amino]-2-methylpropyl}phenyl)acetamide

Preparation 27 (0.075 mmol) was dissolved in ethanol (4 ml) and thesolution treated with a solution of ammonium fluoride (16 mg, 0.43 mmol)in water (300 μL). The reaction mixture was then stirred at 50° C. for18 hours before being allowed to cool to room temperature. The reactionmixture was filtered and washed with methanol:water (2 ml, 1:1 byvolume) to give the title compound as a colourless solid.

¹HNMR (400 MHz, CD₃OD) δ: 1.05-1.16 (m, 9H), 2.70-2.96 (m, 7H), 3.32 and3.34 (2t, 2H), 3.74 and 3.83 (2s, 2H), 4.56 and 4.58 (2s, 2H), 4.64-4.66(m, 1H), 6.85 (dd, 1H), 7.01-7.26 (m, 7H), 7.36 (dd, 1H).

MS (electrospray) m/z 637 [M−H]⁻

Examples 11 to 32

No Q¹ Data 11

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.85 (s, 3 H),0.86 (s, 3 H), 2.47 (s, 3H), 2.60-2.63 (m, 2 H),2.84 (s, 2 H), 2.88 (s, 3 H), 3.61 and 3.75 (2s,2 H), 4.38-4.41 (m, 1 H), 4.89 and 5.03 (2 s, 2 H),6.80 (d, 1 H),6.93-7.18 (m, 9 H), 7.28 and 7.33(2 t, 1 H), 7.59 (d, 1 H), 7.92 and7.74 (2 d, 1 H).MS (electrospray) m/z 604 (M − H]⁻ 12

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.88 (s, 3 H),0.90 (s, 3 H), 2.53 (s, 2H), 2.61-2.63 (m, 2 H),2.88 (s, 3 H), 3.42 (s, 2 H), 4.28 (d, 1 H),4.40-4.43(m, 2 H), 6.82 (t, 1 H), 6.85 (s, 1 H), 6.91 (d,1 H), 6.95 (d,1 H), 7.00 (d, 1 H), 7.05 (s, 1 H),7.07-7.15 (m, 4 H), 7.17 (d, 1 H),7.28 (t, 1 H),7.36-7.40 (m, 2 H) ppm.MS (electrospray) m/z 616 [M − H]⁻13

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.89-0.93 (m,6 H), 2.51-2.54 (m, 2 H,partially obscured bysolvent), 2.58 (s, 2 H), 2.64-2.69 (m, 2 H),2.92(s, 3 H), 3.62-3.66 (m, 2 H), 3.75 (s, 2 H), 4.43-4.46(m, 1 H), 4.50and 4.56 (2 s, 2 H), 6.51-6.60(m, 2 H), 6.73-7.21 (m, 8 H) ppm.MS(electrospray) m/z 568 [M + H]⁺ 14

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.90 (s, 3 H),0.96 (s, 3 H), 1.30-1.50 (m,4 H), 2.40 (t, 2 H),2.46 (s, 2 H), 2.60-2.65 (m, 2 H), 3.89 (s, 3H),2.95-3.08 (m, 2 H), 3.25-3.30 (m, 2 H), 4.40-4.44(m, 1 H), 6.62 (d, 2H), 6.81 (d, 2 H), 6.91 (d, 2 H),6.90-7.06 (m, 4 H), 7.15 (t, 1 H), 7.18(s, 1 H),7.94 (t, 1 H).MS (electrospray) m/z 582 [M − H]⁻,584 [M + H]⁺15

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.90 (s, 3 H),0.96 (s, 3 H), 2.48-2.64 (m,4 H), 2.87 (s, 3 H),3.17 (q, 2 H), 3.30 (s, 2 H), 4.40-4.43 (m, 1H),6.61 (d, 2 H), 6.80 (d, 1 H), 6.85 (d, 2 H), 6.90-7.05(m, 4 H), 7.16(t, 1 H), 7.18 (s, 1 H), 7.98 (t,1 H).MS (electrospray) m/z 554 [M − H]⁻16

¹HNMR (400 MHZ, CD₃OD) δ: 1.06 (s, 3 H), 1.08(s, 3 H), 2.68-2.92 (m, 7H), 3.51 (s, 2 H), 4.34 (s,2 H), 4.64-4.66 (m, 1 H), 6.63 (d, 1 H),6.78-6.79(m, 1 H), 6.86 (d, 1 H), 7.03-7.22 (m, 6 H),7.36-7.37 (m, 1H).MS (electrospray) m/z 574 [M − H]⁻, 576[M + H]⁺, 598 [M + Na]⁺ 17

¹HNMR (400 MHz, CD₃OD) δ: 1.12 (s, 3 H), 1.13(s, 3 H), 2.79 (s, 2 H),2.92 (s, 3 H), 3.00 (d, 2 H),3.51 (s, 2 H), 4.18 (s, 2 H), 4.77-4.80 (m,1 H),6.89 (d, 1 H), 6.97 (s, 1 H), 7.05-7.18 (m, 5 H),7.22-7.26 (m, 1H), 7.40 (s, 1 H).MS (electrospray) m/z 610/612 [M + H]⁺, 608/610[M −H]⁻ 18

¹HNMR (400 MHz, CD₃OD) δ: 1.12 (s, 3 H), 1.13(s, 3 H), 2.77-2.81 (m, 2H), 2.90 (s, 3 H), 2.91-2.97(m, 2 H), 3.53 (s, 2 H), 4.36 (s, 2 H),4.69-4.72(m, 1 H), 6.74 (d, 1 H), 6.88 (d, 1 H), 7.01-7.06(m, 2 H),7.10-7.12 (m, 2 H), 7.16-7.25 (m,2 H), 7.38-7.39 (m, 1 H)MS(electrospray) m/z 610/612 [M + H]⁺, 632/634[M + Na]⁺, 608/610 [M − H]⁻19

¹HNMR (400 MHz, DMSO_(d6)) δ: 1.12 (s, 6 H),2.91-2.98 (m, 7 H), 3.43 (s,2 H), 4.56 (s, 2 H),4.75-4.78 (m, 1 H), 6.76 (d, 1 H), 6.91 (d, 1H),7.04 (d, 1 H), 7.09-7.24 (m, 5 H), 7.30 (s, 1 H),7.39-7.45 (m, 2 H),7.85-7.87 (m, 1 H), 8.13-8.15(m, 1 H) ppm.MS (electrospray) m/z 592 [M +H]⁺, 614[M + Na]⁺, 590 [M − H]⁻ 20

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.93 (s, 3 H),0.95 (s, 3 H), 2.58 (s, 2H), 2.65-2.66 (m, 2 H),2.91 (s, 3 H), 3.46 (s, 2 H), 4.26-4.27 (m, 2H),4.42-4.46 (m, 1 H), 6.82 (d, 1 H), 6.89-6.93 (m,2 H), 6.96-7.01 (m, 3H), 7.05 (s, 1 H), 7.06-7.10(m, 1 H), 7.15-7.19 (m, 2 H) ppm.MS(electrospray) m/z 610 [M + H]⁺, 632[M + Na]⁺, 608 [M − H]⁻ 21

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.88−0.91 (m,6 H), 0.95-0.98 (m, 3 H),2.50-2.63 (m, 4 H), 2.86(s, 3 H), 3.23-3.28 (m, 2 H), 3.58 and 3.72 (2s,2 H), 4.39 and 4.47 (2 s, 3 H), 6.54 (s, 1 H), 6.61-6.67(m, 2 H), 6.78(d, 1 H), 6.92-7.02 (m, 3 H),7.08 (2, 1 H), 7.13-7.15 (m, 2 H) ppm.MS(electrospray) m/z 604 [M + H]⁺, 626 [M + Na]⁺ 22

¹H NMR (400 MHz, CD₃OD): rotamers δ 1.16-1.25(m, 6 H), 2.85-3.10 (m, 10H), 3.79 and 3.84(2 s, 2 H), 4.62 and 4.63 (2 s, 2 H), 4.78-4.82 (m,1H), 6.65-6.70 (m, 2 H), 6.80-6.84 (m, 2 H), 6.88(d, 1 H), 7.01 (d, 1 H),7.08-7.31 (m, 4 H).LRMS (electrospray): m/z [M + H]⁺ 590, [M + Na]⁺612.23

¹H NMR (400 MHz, DMSO_(d6)) δ: (rotamers)0.86-0.90 (m, 6 H), 2.51 and2.55 (2 s, 2 H), 2.58-2.65(m, 2 H), 2.87 (s, 3 H), 2.77 and 2.90 (2 s,3H), 3.66 and 3.73 (2 s, 2 H), 4.38-4.42 (m, 1 H),4.48 and 4.61 (2 s, 2H), 6.77-6.80 (m, 2 H), 6.89-6.91(m, 2 H), 6.93-6.99 (m, 3 H), 7.03-7.06(m,1 H), 7.13-7.18 (m, 2 H) ppm.LRMS (electrospray) : m/z 624 [M + H]⁺.24

¹H NMR (400 MHz, CD₃OD): δ = 1.32 (s, 6 H),2.97 (s, 3 H), 2.99 (s, 2 H),3.11-3.23 (m, 2 H),3.54 (s, 2 H), 4.29-4.31 (m, 2 H), 4.82-4.86 (m,1 H),6.89-6.92 (m, 2 H), 6.95 (d, 1 H), 7.08 (dd,1 H), 7.17-7.24 (m, 6 H),7.29-7.35 (m, 4 H), 7.46(d, 1 H) ppm.LRMS (electrospray): m/z [M + H]⁺618, [M + Na]⁺640, [M − H]⁻ 616. 25

¹H NMR (400 MHz, CD₃OD): δ = 1.33 (s, 3 H),1.34 (s, 3 H), 2.98 (s, 3 H),3.01 (m, 2 H), 3.21-3.25(m, 2 H), 3.56 (s, 2 H), 4.35 (s, 2 H),4.83-4.86(dd, 1 H), 6.75-6.81 (m, 3 H), 6.95 (d, 1 H),7.18-7.25 (m, 6H), 7.28-7.36 (m, 4 H), 7.45 (d,1 H) ppm.LRMS (Electrospray): m/z [M +H]⁺ 618, [M + Na]⁺640. 26

¹H NMR (400 MHz, DMSO_(d6)) δ: 0.91 (s, 3 H),0.93 (s, 3 H), 2.08 (s, 6H), 2.57 (s, 2 H), 2.65-2.66(m, 2 H), 2.89 (s, 3 H), 3.38 (s, 2 H),4.06-4.07(m, 2 H), 4.33-4.46 (m, 1 H), 6.71 (s, 2 H),6.81 (d, 1 H), 6.97(d, 1 H), 7.01 (d, 1 H), 7.05-7.09(m, 2 H), 7.13-7.19 (m, 2 H),8.28-8.31 (m,1 H) ppm.LRMS (APCl): m/z 570 [M + H]⁺ 27

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.90 (m, 6 H),2.41-2.72 (m, 6 H), 2.91 (s,3 H), 3.60 (m, 2 H),3.75 (s, 2 H), 4.40-4.60 (m, 3 H), 6.41-6.59 (m,2H), 6.81-7.18 (m, 8 H) ppm.MS (electrospray) m/z 590 [M + Na]⁺, 566 [M −Na]⁻ 28

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.9 (m, 6 H),2.41 -2.46 (2 H, m),2.54-2.58 (2 H, m), 2.60-2.67(2 H, m), 2.85 (s, 3 H). 3.61 (m, 2 H),3.70 (m,2 H), 4.38-4.41 (m, 1 H), 4.61 (m, 2 H), 6.50-6.61(m, 2 H), 6.81(d, 1 H), 6.91-7.23 (m, 7 H) ppm.MS (electrospray) m/z 568 [M + H]⁺, 590[M + Na]⁺ 29

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.81 (s, 3 H), 0.85(s, 3 H), 2.58 (m, 2H), 2.62 (m, 2 H), 2.89 (s,3 H), 3.41 (d, 1 H), 3.45 (d, 1 H), 3.58 (d,2 H),4.40 (m, 1 H), 4.80 (m, 1 H), 6.80-7.20 (m, 11 H),8.40 (d, 1 H)ppm.MS (APCl) m/z 578 [M + Na]⁺ 30

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.81 (s, 3 H), 0.84(s, 3 H), 2.50 (m, 2H), 2.63 (m, 2 H), 2.85 (s,3 H), 3.40 (m, 2 H), 3.57 (m, 2 H), 4.40 (m,1 H),4.80 (m, 1 H), 6.80 (d, 1 H), 6.98 (m, 1 H), 7.00(m, 9 H), 8.40 (d,1 H) ppm.MS (APCl) m/z 554 [M − H]⁻ 31

¹H NMR (400 MHz, CD₃OD): δ 1.25 (s, 3 H), 1.26(s, 3 H), 2.93 (s, 2 H),2.96 (s, 3 H), 3.07-3.09 (m,2 H), 3.62 (s, 2 H), 4.44 (s, 2 H),4.77-4.80 (m,1 H), 6.77-6.79 (m, 1 H), 6.93 (d, 1 H), 7.01-7.02(m, 1 H),7.05-7.07 (m, 1 H), 7.14-7.18 (m, 2 H),7.20-7.27 (m, 3 H), 7.29-7.34 (m,3 H), 7.43 (d,1 H), 7.51 (s, 1 H), 7.54 (s, 1 H) ppm.LRMS(Electrospray): m/z [M + H]⁺ 618, [M + Na]⁺640. 32

¹H NMR (400 MHz, CD₃OD): δ 1.14 (s, 3 h), 1.16(s, 3 H), 2.77-3.00 (m, 7H), 3.60 (s, 2 H), 4.44 (s,2 H), 4.69-4.72 (dd, 1 H), 6.89-6.93 (m, 3H),7.09-7.30 (m, 9 H), 7.43 (d, 1 H), 7.50 (s, 1 H),7.52 (s, 1 H)ppm.LRMS (Electrospray): m/z [M − H]⁻ 616 Examples 34-38

33

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.95 (s, 3 H), 0.98(s, 3 H), 2.65-2.71 (m,4 H), 2.93 (s, 3 H), 4.46-4.52(m, 3 H), 6.83-6.86 (m, 3 H), 7.03-7.06(m,1 H), 7.22-7.23 (m, 1 H), 7.30-7.38 (m, 4 H), 7.47(d, 2 H), 7.54 (d,2 H), 7.73-7.76 (m, 2 H),8.96-8.99 (m, 1 H) ppm.MS (electrospray) m/z604 [M + H]⁺ 34

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.95 (s, 3 H), 0.97(s, 3 H), 1.26 (s, 6H), 2.63-2.70 (m, 4 H), 2.93 (s,3 H), 3.39-3.40 (m, 2 H), 4.44-4.48 (m,1 H), 6.71(d, 2 H), 6.84 (d, 1 H), 7.05 (d, 1 H), 7.20-7.34 (m,4 H),7.58-7.60 (m, 2 H), 8.02-8.06 (m, 1 H) ppm.MS (electrospray) m/z 570[M + H]⁺ 35

¹HNMR (400 MHz, DMSO_(d6)) δ: 0.95 (s, 3 H), 0.98(s, 3 H), 2.64-2.71 (m,4 H), 2.93 (s, 3 H), 4.45-4.48(m, 1 H), 4.54-4.56 (m, 2 H), 6.82-6.86(m,3 H), 7.04 (d, 1 H), 7.22-7.40 (m, 5 H), 7.45-7.48(m, 3 H), 7.54 (s,1 H), 7.73-7.75 (m, 2 H),8.98-9.01 (m, 1 H) ppm.MS (electrospray) m/z604 [M + H]⁺ 36

¹HNMR (400 MHz, CD₃OD) δ: 1.05 (s, 3 H), 1.12(s, 3 H), 2.08 (s, 3 H),2.22 (s, 3 H), 2.70-2.97 (m,9 H), 3.46-3.50 (m, 2 H), 4.63-4.66 (m, 1H), 6.54(s, 1 H), 6.86 (d, 1 H), 6.88 (s, 1 H), 7.09-7.11 (m,1 H),7.30-7.38 (m, 3 H), 7.61 (m, 1 H), 7.66 (d,1 H) ppm.MS (electrospray)m/z 570 [M + H]⁺ 37

¹HNMR (400 MHz, CD₃OD) δ: 1.05 (s, 3 H), 1.12(s, 3 H), 2.11 (s, 3 H),2.23 (s, 3 H), 2.70-2.97 (m,9 H), 3.45-3.49 (m, 2 H), 4.63-4.66 (m, 1H), 6.54(d, 1 H), 6.81 (d, 1 H), 6.87 (d, 1 H), 7.09-7.12 (m,1 H),7.30-7.38 (m, 3 H), 7.61 (m, 1 H), 7.66 (d,1 H) ppm.MS (electrospray)m/z 570 [M + H]⁺, 569 [M − H]⁻ 38

¹HNMR (400 MHz, CD₃OD) δ: 1.04 (s, 3 H), 1.11(s, 3 H), 2.12 (s, 3 H),2.70-2.96 (m, 9 H), 3.51-3.54(m, 2 H), 4.63-4.66 (m, 1 H), 6.64 (d, 1H),6.86 (d, 2 H), 6.94 (m, 1 H), 7.09 (d, 1 H), 7.30-7.38(m, 3 H), 7.59(m, 1 H), 7.64 (d, 1 H) ppm.MS (electrospray) m/z 556 [M + H]⁺ Unlessotherwise stated all reactions were carried out under a nitrogenatmosphere. Abbreviations TBDMS = tert-butyl(dimethyl)silyl IPA:isopropyl alcohol THF: Tetrahydrofuran s = singlet d = doublet dd =double doublet t = triplet q = quartet m = multiplet bs = broad singlete.g. NH, or OH

The ability of the compounds of the formula (1) to act as potent β2agonists therefore mediating smooth muscle relaxation may be determinedby the measure of the effect of beta-2 adrenergic receptor stimulationon electrical field stimulated-contraction of guinea pig trachea strips.

Guinea-Pig Trachea

Male, Dunkin-Hartley guinea pigs (475-525 g) are killed by CO₂asphyxiation and exsanguination from the femoral artery and the tracheais isolated. Four preparations are obtained from each animal, startingthe dissection immediately below the larynx and taking 2.5 cm length oftrachea. The piece of trachea is opened by cutting the cartilageopposite the trachealis muscle, then transverse sections, 34 cartilagerings wide, are cut. The resulting strip preparations are suspended in 5ml organ baths using cotton threads tied through the upper and lowercartilage bands. The strips are equilibrated, un-tensioned, for 20minutes in a modified Krebs Ringer buffer (Sigma K0507) containing 3 μMIndomethacin (Sigma I7378), 10 μM Guanethidine (Sigma G8520) and 10 μMAtenolol (Sigma A7655), heated at 37° C. and gassed with 95% O₂/5% CO₂,before applying an initial tension of 1 g. The preparations are allowedto equilibrate for a further 30-45 minutes, during which time they arere-tensioned (to 1 g) twice at 15-minute intervals. Changes in tensionare recorded and monitored via standard isometric transducers coupled toa data-collection system (custom-designed at Pfizer). Following thetensioning equilibration, the tissues are subjected to electrical fieldstimulation (EFS) using the following parameters: 10 s trains every 2minutes, 0.1 ms pulse width, 10 Hz and just-maximal voltage (25 Volts)continuously throughout the length of the experiment. EFS ofpost-ganglionic cholinergic nerves in the trachea results in monophasiccontractions of the smooth muscle and twitch height is recorded. Theorgan baths are constantly perfused with the above-described KrebsRinger buffer by means of a peristaltic pump system (pump flow rate 7.5ml/minute) throughout the experiment, with the exception of when abeta-2 agonist according to the present invention is added, the pump isthen stopped for the time of the cumulative dosing to the bath andstarted again after maximal response is reached for the wash-out period.

Experimental Protocol for Assessment of Potency and Efficacy

Following equilibration to EFS, the peristaltic pump is stopped and thepreparations ‘primed’ with a single dose of 300 nM isoprenaline (SigmaI5627) to establish a maximal response in terms of inhibition of thecontractile EFS response. The isoprenaline is then washed out over aperiod of 40 minutes. Following the priming and wash-out recovery, astandard curve to isoprenaline is carried out on all tissues(Isoprenaline Curve 1) by means of cumulative, bolus addition to thebath using half log increments in concentration. The concentration rangeused is 1^(e-9) to 1^(e)/3^(e-6)M. At the end of the isoprenaline curvethe preparations are washed again for 40 minutes before commencing asecond curve, either to isoprenaline (as internal control) or a beta-2agonist according to the present invention. Beta-2 agonist responses areexpressed as percentage inhibition of the EFS response. Data for beta-2agonist are normalised by expressing inhibition as a percentage of themaximal inhibition induced by isoprenaline in Curve 1. The EC₅₀ valuefor beta-2 agonist according to the present invention refers to theconcentration of compound required to produce half maximal effect. Datafor beta-2 agonists according to the present invention are thenexpressed as relative potency to isoprenaline defined by the ratio (EC₅₀beta-2 agonist)/(EC50 Isoprenaline).

Confirmation of Beta-2 Mediated Functional Activity

Beta-2 agonist activity of test compounds is confirmed using theprotocol above, however, prior to constructing the curve to beta-2agonist according to the present invention, the preparations arepre-incubated (for a minimum of 45 minutes) with 300 nM ICI 118551 (aselective β₂ antagonist) which results in the case of a beta-2 mediatedeffect in a rightward-shift of the test compound dose response curve.

According to another alternative, the agonist potency for the β2receptor of the compounds of the formula (1) may also be determined bythe measure of the concentration of compound according to the presentinvention required to produce half maximal effect (EC₅₀) for the β2receptor.

Compound Preparation

10 mM/100% DMSO (dimethylsulfoxide) stock of compound is diluted torequired top dose in 4% DMSO. This top dose is used to construct a10-point semi-log dilution curve, all in 4% DMSO. Isoprenaline (Sigma,1-5627) was used as a standard in every experiment and for control wellson each plate. Data was expressed as % Isoprenaline response.

Cell Culture

CHO (Chinese Hamster Ovary) cells recombinantly expressing the human β2adrenergic receptor (from Kobilka et al., PNAS 84: 46-50, 1987 andBouvier et al., Mol Pharmacol 33: 133-139 1988 CHOhβ2) were grown inDulbeccos MEM/NUT MIX F12 (Gibco, 21331-020) supplemented with 10%foetal bovine serum (Sigma, F4135, Lot 90K8404 Exp 09/04), 2 mMglutamine (Sigma, G7513), 500 μg/ml geneticin (Sigma, G7034) and 10μg/ml puromycin (Sigma, P8833). Cells were seeded to give about 90%confluency for testing.

Assay Method

25 μl/well each dose of compound was transferred into a cAMP-Flashplate®(NEN, SMP004B), with 1% DMSO as basal controls and 100 nM Isoprenalineas max controls. This was diluted 1:2 by the addition of 25 μl/well PBS.Cells were trypsinised (0.25% Sigma, T4049), washed with PBS (Gibco,14040-174) and resuspended in stimulation buffer (NEN, SMP004B) to give1×10⁶ cells/ml CHOhB2. Compounds were incubated with 50 μl/well cellsfor 1 hour. Cells were then lysed by the addition of 100 μl/welldetection buffer (NEN, SMP004B) containing 0.18 μCi/ml ¹²⁵I-cAMP (NEN,NEX-130) and plates were incubated at room temperature for a further 2hours. The amount of ¹²⁵I-cAMP bound to the Flashplate® was quantifiedusing a Topcount NXT (Packard), normal counting efficiency for 1 minute.Dose-response data was expressed as % Isoprenaline activity and fittedusing a four parameter sigmoid fit. It has thus been found that thecompounds of formula (1) according to the present invention that areillustrated in examples 1 to 38 above show a β2 CAMP EC₅₀ between 0.02nM and 3.03 nM. The following table illustrate the activity of thecompounds of the invention:

Example EC₅₀ (nM) 1 0.03 2 0.24 3 0.02 6 0.16 7 0.26 9 0.45 13 0.08 150.09 20 0.29

1-23. (canceled)
 24. A method of treating an allergic or non-allergicairways disease in a mammal, said method comprising administering tosaid mammal an effective amount of a compound of the formula (1):

or a pharmaceutically acceptable salt, isomer or tautomer thereof,wherein: the (CH₂)_(n)—C(═O)Q¹ group is in the meta or para position; R¹and R² are independently selected from H and C₁-C₄ alkyl; n is 0, 1 or2; and Q¹ is selected from:

and *—NR⁸-Q²-A; p is 1 or 2; Q² is C₁-C₄ alkylenyl optionallysubstituted with one hydroxy group; R⁸ is H or C₁-C₄ alkyl; A is pyridyloptionally substituted with OH, C₃-C₇ cycloalkyl optionally substitutedwith OH, or

R³, R⁴, R⁵, R⁶ and R⁷ are independently H, C₁-C₄ alkyl OR⁹, SR⁹, halo,CN, CF₃, OCF₃, COOR⁹, SO₂NR⁹R¹⁰, CONR⁹R¹⁰, NR⁹R¹⁰, NHCOR¹⁰ or phenyloptionally substituted with 1 to 3 OR⁹, halo or C₁-C₄ alkyl; R⁹ and R¹⁰are independently H or C₁-C₄ alkyl; and the * represents the attachmentpoint to the carbonyl group of formula (1); provided that the group Q′is substituted with at least one hydroxy group, or a pharmaceuticalcomposition comprising said compound of formula (1), an isomer ortautomer thereof, or a pharmaceutically acceptable salt of saidcompound, isomer or tautomer, and a pharmaceutically acceptableexcipient or additive.
 25. A method of claim 24 wherein said allergic ornon-allergic airways disease is selected from the group consisting of:asthma, acute bronchoconstruction, bronchitis, small airwaysobstruction, emphysema, chronic eosinophilic pneumonia, adultrespiratory distress syndrome (ARDS), acute lung injury andbronchiectasis.
 26. A method of claim 25 wherein said asthma is selectedfrom the group consisting of: atopic asthma, non-atopic asthma, allergicasthma, atopic bronchial IgE-mediated asthma, bronchial asthma,essential asthma, true asthma, intrinsic asthma caused bypathophysiologic disturbances, extrinsic asthma caused by environmentalfactors, essential asthma of unknown or inapparent cause, bronchiticasthma, emphysematous asthma, exercise-induced asthma, allergen inducedasthma, cold air induced asthma, occupational asthma, infective asthmacaused by bacterial, fungal, protozoal, or viral infection, non-allergicasthma, incipient asthma, wheezy infant syndrome and bronchiolytis. 27.A method of claim 25 wherein said bronchitis is selected from the groupconsisting of acute bronchitis, chronic bronchitis, acutelaryngotracheal bronchitis, arachidic bronchitis, catarrhal bronchitis,croupus bronchitis, dry bronchitis, infectious asthmatic bronchitis,productive bronchitis, staphylococcus bronchitis, streptococcalbronchitis and vesicular bronchitis.
 28. A method of claim 25 whereinsaid bronchiectasis is selected from the group consisting of cylindricbronchiectasis, sacculated bronchiectasis, fusiform bronchiectasis,capillary bronchiectasis, cystic bronchiectasis, dry bronchiectasis andfollicular bronchiectasis.
 29. A pharmaceutical composition comprising acombination of a compound of formula (1),

wherein: the (CH₂)_(n)—C(═O)Q¹ group is in the meta or para position; R¹and R² are independently selected from H and C₁-C₄ alkyl; n is 0, 1 or2; and Q¹ is selected from:

and *—NR⁸-Q²-A; p is 1 or 2; Q² is C₁-C₄ alkylenyl optionallysubstituted with one hydroxy group; R⁸ is H or C₁-C₄ alkyl; A is pyridyloptionally substituted with OH, C₃-C₇ cycloalkyl optionally substitutedwith OH, or

R³, R⁴, R⁵, R⁶ and R⁷ are independently H, C₁-C₄ alkyl, OR⁹, SR⁹, halo,CN, CF₃, OCF₃, COOR⁹, SO₂NR⁹R¹⁰, CONR⁹R¹⁰, NR⁹R¹⁰, NHCOR¹⁰ or phenyloptionally substituted with 1 to 3 OR⁹, halo or C₁-C₄ alkyl; R⁹ and R¹⁰are independently H or C₁-C₄ alkyl; and the * represents the attachmentpoint to the carbonyl group of formula (1); provided that the group Q¹is substituted with at least one hydroxy group,

or a pharmaceutically acceptable salt, isomer or tautomer thereof, witha 5-lipoxygenase (5-LO) inhibitor or a 5-lipoxygenase activating protein(FLAP) antagonist; a leukotriene (LTRA) antagonist; an LTB₄, LTC₄, LTD₄,or LTE₄ antagonist; a histamine receptor antagonist; a H1 or H3antagonist; an α₁- and α₂-adrenoceptor agonist vasoconstrictorsympathomimetic agents for decongestant use; a muscarinic M3 receptorantagonist or anticholinergic agent; a PDE3, PDE4 or PDE5 inhibitor;theophylline; sodium cromoglycate; a non-selective or selective COX-1 orCOX-2 inhibitor; an oral and inhaled glucocorticosteroids; a monoclonalantibody active against endogenous inflammatory entities; an anti-tumornecrosis factor; an anti-TNFα agent; an adhesion molecule inhibitor; aVLA-4 antagonists; a kinin-B₁- or B₂-receptor antagonist; animmunosuppressive agents; an inhibitors of matrix metalloprotease; atachykinin NK₁, NK₂ or NK₃ receptor antagonist; an elastase inhibitor;an adenosine A2a receptor agonist; an inhibitors of urokinase; acompound that acts on dopamine receptors; a D2 agonist; a modulators ofthe NFκβpathway; an IKK inhibitor; a modulator of cytokine signallingpathways; a p38 MAP kinase or syk kinase modulator; a mucolytic oranti-tussive agent; or an antibiotic.